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- | '''β班の実験ノート'''
| + | <html><link rel="stylesheet" href="https://2008.igem.org/wiki/index.php?title=User:Maiko/chiba.css&action=raw&ctype=text/css" type="text/css" /></html> |
| + | >[[Team:Chiba/Internal|ノートに戻る]] |
| | | |
- | ==Week 1==
| |
| | | |
- | ===17 August, 2008===
| |
- | :'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| |
- | :competent cells : XL10G
| |
- | ::*BBa_R0040(2008) [http://partsregistry.org/Part:BBa_R0040] -> No colonies on plates
| |
- | ::*BBa_C0070(2008) [http://partsregistry.org/Part:BBa_C0070] -> No colonies on plates
| |
- | ::*BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates
| |
- | ::*BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] -> No colonies on plates
| |
- | ::*BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] -> No colonies on plates
| |
- | ::*BBa_C0079(2008) [http://partsregistry.org/Part:BBa_C0079] -> No colonies on plates
| |
| | | |
- | ===18 August, 2008=== | + | ==送受信班の実験ノート== |
- | :'''[[Team:Chiba/protocol/PCR|PCR]]'''
| + | *[[Team:Chiba/jk/β/week 1|week 1]] |
- | ::*BBa_C0070(2008) [http://partsregistry.org/Part:BBa_C0070]
| + | *[[Team:Chiba/jk/β/week 2|week 2]] |
- | ::*BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076]
| + | *[[Team:Chiba/jk/β/week 3|week 3]] |
- | ::*BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077]
| + | *[[Team:Chiba/jk/β/week 4|week 4]] |
- | ::*BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078]
| + | *[[Team:Chiba/jk/β/week 5|week 5]] |
- | ::*BBa_C0079(2008) [http://partsregistry.org/Part:BBa_C0079]
| + | *[[Team:Chiba/jk/β/week 6|week 6]] |
- | ::*BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]・・・control
| + | |
| | | |
| + | ==モジュール解説== |
| + | ===感度差による応答時間の差=== |
| + | 任意の順番と、delay-timeをもった遺伝子発現を実現するために、バクテリア間の細胞間通信、クオラムセンシングを利用する。クオラムセンシングでは、応答閾値を超えるAHLが蓄積されたとき初めて、目的遺伝子の発現がおこる。AHL情報がじゅうぶんゆっくり蓄積するとき、ひとつのAHL送信装置に対して、感度の異なる複数の受信装置によってさまざまな出力装置を独立に起動すれば、感度の高いAHL 受信機ほど先に起動する。 |
| | | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | === Controlling the Delay time of the AHL signaling === |
- | <tr>
| + | LuxIのつくるAHLに対して、感度差による応答時間差は、2つの方法で実現することができる。 |
- | <td width="257">DNA Template</td>
| + | #LuxR/ Pluxの感度変異体を用いる場合<br>LuxRの変異体を用いれば、pluxがonになるAHL濃度が<1nMから~10nMまで変化する。[http://authors.library.caltech.edu/5553/ C. H. Collins.''et al.:''Mol.Microbiol.2005.'''55'''(3).712–723] |
- | <td>1</td>
| + | #クロストークを使う場合 |
- | </tr>
| + | :AHL=Acyl-homoserine Lactoneには、そのアシル基が異なる種類がある。luxIが合成するAHLは、3-oxo-hyxanoyl-Homoserine Lactone (=OHHL)であり、3-oxo-dodecanoyl-homoserine Lactone(=ODHL)や、butyryl-homoserine Lactone(=HHL)が存在する。それぞれ、合成酵素はLasI、RhlI(ともにシュードモナス由来)と呼ばれる。このほかのAHL分子と、その合成酵素を以下に示した。これらのAHL分子は、アシル基のみが異なるため、一定の割合でLuxRと相互作用する(cross-talkする)。たとえば、OdHLとLuxRとのcross-talk(LasI-LuxR)は、OHHLとLuxRの場合(LuxI-LuxR)の1/20である。AHL分子の蓄積速度が一定ならば,luxI-LuxR/Pluxよりも20倍遅くスイッチが入ることとなる。 |
- | <tr> | + | |
- | <td>dNTP mix</td>
| + | |
- | <td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Foward Primer</td>
| + | |
- | <td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Reverse Primer</td>
| + | |
- | <td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>DNA polymerase</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Thermopol Buffer</td>
| + | |
- | <td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>68</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>100μL</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | :95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃ | + | |
- | | + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | | + | |
| | | |
- | {|align="justify"
| + | *AHLsおよびその合成酵素 |
- | |[[Image:Chiba-0818.JPG]]
| + | <table width="500" border="1" cellpadding="0" cellspacing="0" bordercolor="#000000" center> |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
| <tr> | | <tr> |
- | <td width="257">Sample DNA</td> | + | <td width="200">Strain</td> |
- | <td>3</td> | + | <td width="200">Signal Molecule</td> |
| + | <td width="150">Enzyme<td> |
| </tr> | | </tr> |
| <tr> | | <tr> |
- | <td>Loading Dye</td> | + | <td>P.aeruginosa</td> |
- | <td>2</td> | + | <td>C4-HSL</td> |
| + | <td>RhlI</td> |
| </tr> | | </tr> |
| <tr> | | <tr> |
- | <td>dH2O</td> | + | <td>V. fisheri</td> |
- | <td>7</td> | + | <td>C6-3-oxo-HSL</td> |
| + | <td>LuxI</td> |
| </tr> | | </tr> |
| <tr> | | <tr> |
- | <td>TOTAL</td> | + | <td>A.tumefaciens</td> |
- | <td>12μL</td> | + | <td>C8-3-oxo-HSL</td> |
| + | <td>TraI</td> |
| </tr> | | </tr> |
- | </table>
| |
- |
| |
- | :From left;
| |
- | ::*BBa_C0070(2008) [http://partsregistry.org/Part:BBa_C0070] ->None
| |
- | ::*BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] ->None
| |
- | ::*BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] ->None
| |
- | ::*BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] ->None
| |
- | ::*BBa_C0079(2008) [http://partsregistry.org/Part:BBa_C0079] ->None
| |
- | ::*BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]・・・control ->OK
| |
- | |}
| |
- |
| |
- |
| |
- |
| |
- |
| |
- | :'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| |
- | competent cells : XL10G
| |
- |
| |
- | ::*BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
| |
- | ::*BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
| |
- | ::*BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
| |
- | ::*BBa_C0076(2006) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates(19/8)
| |
- | ::*BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
| |
- | ::*BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
| |
- | ::*BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
| |
- | ::*BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
| |
- | ::*BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
| |
- | ::*BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]
| |
- |
| |
- |
| |
- | :--->(20/8)[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]
| |
- |
| |
- | ===19 August, 2008===
| |
- | :'''[[Team:Chiba/protocol/PCR|PCR]]'''
| |
- | ::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
| |
- | ::*BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
| |
- | ::*BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
| |
- | ::*BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
| |
- | ::*BBa_S3154(2007) [http://partsregistry.org/Part:BBa_S3154]
| |
- | ::*BBa_S3154(2006) [http://partsregistry.org/Part:BBa_S3154]
| |
- | ::*BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]・・・control
| |
- |
| |
- |
| |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| <tr> | | <tr> |
- | <td width="257">DNA Template</td> | + | <td>P.aeruginosa</td> |
- | <td>1</td> | + | <td>C10-3-oxo-HSL</td> |
| + | <td>VanI</td> |
| </tr> | | </tr> |
- | <tr>
| |
- | <td>dNTP mix</td>
| |
- | <td>10</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>Foward Primer</td>
| |
- | <td>5</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>Reverse Primer</td>
| |
- | <td>5</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>DNA polymerase VENT</td>
| |
- | <td>1</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>Thermopol Buffer</td>
| |
- | <td>5</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>dH2O</td>
| |
- | <td>28</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>TOTAL</td>
| |
- | <td>50μL</td>
| |
- | </tr>
| |
- | </table>
| |
| | | |
- |
| |
- | :95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃
| |
- |
| |
- |
| |
- |
| |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
- | {|align="justify"
| |
- | |[[Image:Chiba-0819.JPG]]
| |
- | |
| |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| <tr> | | <tr> |
- | <td width="257">Sample DNA</td> | + | <td>P.aeruginosa</td> |
- | <td>3</td> | + | <td>C12-3-oxo-HSL</td> |
| + | <td>LasI</td> |
| </tr> | | </tr> |
- | <tr>
| |
- | <td>Loading Dye</td>
| |
- | <td>2</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>dH2O</td>
| |
- | <td>7</td>
| |
- | </tr>
| |
- | <tr>
| |
- | <td>TOTAL</td>
| |
- | <td>12μL</td>
| |
- | </tr>
| |
- | </table>
| |
- |
| |
- | :From left;
| |
- | ::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] ->None
| |
- | ::*BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026] ->None
| |
- | ::*BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030] ->None
| |
- | ::*BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030] ->None
| |
- | ::*BBa_S3154(2007) [http://partsregistry.org/Part:BBa_S3154] ->None
| |
- | ::*BBa_S3154(2006) [http://partsregistry.org/Part:BBa_S3154] ->None
| |
- | ::*BBa_F2620(2007) [http://partsregistry.org/Part:BBa_F2620]・・・control ->OK
| |
- | |}
| |
- |
| |
- |
| |
- |
| |
- | ===20 August, 2008===
| |
- | :(18/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| |
- | ::*BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
| |
- | ::*BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070]
| |
- | ::*BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
| |
- | ::*BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
| |
- | ::*BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
| |
- | ::*BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
| |
- | ::*BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
| |
- | ::*BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
| |
- | ::*BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]
| |
- |
| |
| | | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| |
- | {|align="justify"
| |
- | |[[Image:Chiba-0820-3.JPG]]
| |
- | |
| |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| |
| <tr> | | <tr> |
- | <td width="257">Sample DNA</td> | + | <td>R.leguminosarum</td> |
- | <td>3</td> | + | <td>C14-3-hydroxy-HSL</td> |
- | </tr>
| + | <td>CinI</td> |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>7</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>12μL</td> | + | |
| </tr> | | </tr> |
| </table> | | </table> |
| + | [http://partsregistry.org/Part:BBa_F2620:Specificity AHLsについて] |
| | | |
- | :From left;
| + | ==実験== |
- | ::*BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None
| + | ===Construction(From BioBrick)=== |
- | ::*BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None
| + | :*Sender(Autoinducer synthase) |
- | ::*BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> None
| + | ::LuxI,LasI,CinI,RhlI(lacプロモーター) |
- | |}
| + | :*Receiver |
- | | + | ::LuxR,LasR,CinR,RhlR |
- | | + | |
- | | + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0820-4.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample DNA</td>
| + | |
- | <td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>24μL</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | :From left;
| + | |
- | ::*BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None
| + | |
- | ::*BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> OK
| + | |
- | ::*BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None
| + | |
- | ::*BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070] -> OK
| + | |
- | ::*BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070] -> OK
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | | + | |
- | | + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0820.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample DNA</td>
| + | |
- | <td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>7</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>12μL</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | :From left;<br>
| + | |
- | ::*BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK<br>
| + | |
- | ::*BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK<br>
| + | |
- | ::*BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK<br>
| + | |
- | ::*BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK<br>
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | --->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
| + | |
- | ::*BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]
| + | |
- | ::*BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]
| + | |
- | ::*BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]
| + | |
- | ::*BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]
| + | |
- | | + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1</td><td>2</td><td>3</td><td>4</td><td>5</td><td>6</td><td>7</td><td>8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170]</td>
| + | |
- | <td>3</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170]</td>
| + | |
- | <td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178]</td>
| + | |
- | <td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178]</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>EcoRⅠ</td>
| + | |
- | <td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>PstⅠ</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>-</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer EcoRⅠ</td>
| + | |
- | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td>
| + | |
- | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>4.9</td><td>4.9</td><td>4.9</td><td>4.9</td><td>4.8</td><td>4.8</td><td>4.8</td><td>4.8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0820-2.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample DNA</td>
| + | |
- | <td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>7</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>12μL</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | :From left;
| + | |
- | :(Single Digestion 1~4)
| + | |
- | ::*BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK
| + | |
- | ::*BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK
| + | |
- | ::*BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK
| + | |
- | ::*BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK
| + | |
- | :(Double Digestion 1~4)
| + | |
- | ::*BBa_C0170(2006) [http://partsregistry.org/Part:BBa_C0170] -> OK
| + | |
- | ::*BBa_C0170(2007) [http://partsregistry.org/Part:BBa_C0170] -> OK
| + | |
- | ::*BBa_C0178(2006) [http://partsregistry.org/Part:BBa_C0178] -> OK
| + | |
- | ::*BBa_C0178(2007) [http://partsregistry.org/Part:BBa_C0178] -> OK
| + | |
- | |}
| + | |
- | | + | |
- | ===21 August, 2008===
| + | |
- | :(20/8)--->'''[[Team:Chiba/protocol/PCR|PCR]]'''
| + | |
- | ::*BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070]
| + | |
- | ::*BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070] | + | |
- | ::*BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076]
| + | |
- | ::*BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078]
| + | |
- | ::*BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078]
| + | |
- | | + | |
- | | + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">DNA Template</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dNTP mix</td>
| + | |
- | <td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Foward Primer</td>
| + | |
- | <td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Reverse Primer</td>
| + | |
- | <td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>DNA polymerase TAQ</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Thermopol Buffer</td>
| + | |
- | <td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>28</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>50μL</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | :95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0821.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample DNA</td>
| + | |
- | <td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>7</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>12μL</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | :From left;
| + | |
- | ::*BBa_C0078(2007) [http://partsregistry.org/Part:BBa_C0078] -> None
| + | |
- | ::*BBa_C0078(2006) [http://partsregistry.org/Part:BBa_C0078] -> None
| + | |
- | ::*BBa_C0076(2007) [http://partsregistry.org/Part:BBa_C0076] -> None
| + | |
- | ::*BBa_C0070(2007) [http://partsregistry.org/Part:BBa_C0070] -> OK
| + | |
- | ::*BBa_C0070(2006) [http://partsregistry.org/Part:BBa_C0070] -> OK
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| + | |
- | :competent cells : XL10G
| + | |
- | ::*BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
| + | |
- | ::*BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
| + | |
- | ::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
| + | |
- | ::*BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
| + | |
- | ::*BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
| + | |
- | ::*BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
| + | |
- | | + | |
- | | + | |
- | | + | |
- | | + | |
- | --->(23/8)'''[[Team:Chiba/protocol/digestion|Digestion]]'''
| + | |
- | | + | |
- | | + | |
- | | + | |
- | ===22 August, 2008===
| + | |
- | :'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| + | |
- | :competent cells : XL10G
| + | |
- | ::*BBa_J13002(2007) [http://partsregistry.org/Part:BBa_J13002]
| + | |
- | ::*BBa_J13002(2006) [http://partsregistry.org/Part:BBa_J13002]
| + | |
- | ::*BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]
| + | |
- | ::*BBa_J04500(2006) [http://partsregistry.org/Part:BBa_J04500]
| + | |
- | :(Locate & 2 Punch ~iyama ver.~)
| + | |
- | ::*BBa_C0076(2008) [http://partsregistry.org/Part:BBa_C0076] -> No colonies on plates
| + | |
- | ::*BBa_C0077(2008) [http://partsregistry.org/Part:BBa_C0077] -> No colonies on plates
| + | |
- | ::*BBa_C0078(2008) [http://partsregistry.org/Part:BBa_C0078] -> No colonies on plates
| + | |
- | ::*BBa_J04500(2008) [http://partsregistry.org/Part:BBa_J04500] -> No colonies on plates
| + | |
- | | + | |
- | | + | |
- | | + | |
- | --->(24/8)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| + | |
- | | + | |
- | ===23 August, 2008===
| + | |
- | :(21/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| + | |
- | ::*BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
| + | |
- | ::*BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
| + | |
- | ::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
| + | |
- | ::*BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
| + | |
- | ::*BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
| + | |
- | ::*BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
| + | |
- | ::*BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
| + | |
- | ::*BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
| + | |
- | ::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
| + | |
- | ::*BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
| + | |
- | ::*BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
| + | |
- | ::*BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
| + | |
- | ::*BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]
| + | |
- | | + | |
- | | + | |
- | :<table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="270">Sample No</td>
| + | |
- | <td> 1</td><td> 2</td><td> 3</td><td> 4</td><td> 5</td><td> 6</td><td> 7</td><td> 8</td><td> 9</td><td>10</td><td>11</td><td>12</td><td>13</td><td>14</td><td>15</td><td>16</td><td>17</td><td>18</td><td>19</td><td>20</td><td>21</td>
| + | |
- | </tr><tr>
| + | |
- | <td>BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]</td>
| + | |
- | <td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]</td>
| + | |
- | <td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]</td>
| + | |
- | <td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td></tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td></tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>3</td></tr>
| + | |
- | <tr>
| + | |
- | <td>EcoRⅠ</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>PstⅠ</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer EcoRⅠ</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>0.9</td><td>0.9</td><td>0.9</td><td>0.9</td><td>0.9</td><td>0.9</td><td>0.9</td><td>0.8</td><td>0.8</td><td>0.8</td><td>0.8</td><td>0.8</td><td>0.8</td><td>0.8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>-</td><td>5</td><td>5</td><td>5</td><td>5</td><td>5</td><td>5</td><td>5</td><td>5</td><td>5</td><td>5</td><td>5</td><td>5</td><td>5</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>3</td><td>3</td><td>3</td><td>3</td><td>3</td><td>3</td><td>1</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :--->[[Team:Chiba/protocol/gelcheck|Gel Check]]
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0823-1.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1~6</td><td>7</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>3</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>2</td><td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6</td><td>6</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | :From left; Sanple No. 1~7
| + | |
- | ::*BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
| + | |
- | ::*BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154]
| + | |
- | ::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
| + | |
- | ::*BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026]
| + | |
- | ::*BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
| + | |
- | ::*BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
| + | |
- | ::*BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | | + | |
- | | + | |
- | | + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0823-2.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>8~21</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>12</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | :From left;
| + | |
- | :(Single Digestion) Sanple No. 8~14
| + | |
- | ::*BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154] -> OK
| + | |
- | ::*BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154] -> OK
| + | |
- | ::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] -> OK
| + | |
- | ::*BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026] -> OK
| + | |
- | ::*BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030] -> OK
| + | |
- | ::*BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030] -> OK
| + | |
- | ::*BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500] -> OK
| + | |
- | | + | |
- | :(Double Digestion) Sample No. 15~21
| + | |
- | ::*BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154] -> OK
| + | |
- | ::*BBa_S03154(2006) [http://partsregistry.org/Part:BBa_S03154] -> OK
| + | |
- | ::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026] -> OK
| + | |
- | ::*BBa_I9026(2006) [http://partsregistry.org/Part:BBa_I9026] -> OK
| + | |
- | ::*BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030] -> OK
| + | |
- | ::*BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030] -> OK
| + | |
- | ::*BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500] -> OK?
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | | + | |
- | | + | |
- | :'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
| + | |
- | ::BBa_J04500(2007:old-sample)[http://partsregistry.org/Part:BBa_J04500]
| + | |
- | | + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1</td><td>2</td><td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>3</td><td>3</td><td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>EcoRⅠ</td>
| + | |
- | <td>-</td><td>0.1</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>SpeⅠ</td>
| + | |
- | <td>-</td><td>-</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>EcoRⅠ Buffer</td>
| + | |
- | <td>-</td><td>0.9</td><td>0.8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td>
| + | |
- | <td>-</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>-</td><td>5</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>3</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | --->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0823-3.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1</td><td>2</td><td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>3</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>1</td><td>2</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>2</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6</td><td>12</td><td>12</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | :From left;
| + | |
- | ::#BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500] -> OK
| + | |
- | ::#BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]Single Digestion -> OK
| + | |
- | ::#BBa_J04500(2007) [http://partsregistry.org/Part:BBa_J04500]Double Digestion -> Bad
| + | |
- | |}
| + | |
- | | + | |
- | ==Week 2==
| + | |
- | ===24 August, 2008===
| + | |
- | :(22/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| + | |
- | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
| + | |
- | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
| + | |
- | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
| + | |
- | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
| + | |
- | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
| + | |
- | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)
| + | |
- | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)
| + | |
- | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006)
| + | |
- | | + | |
- | | + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>5</td><td>6</td><td>7</td><td>8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)</td>
| + | |
- | <td>1</td><td>-</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)</td>
| + | |
- | <td>-</td><td>1</td><td>-</td><td>-</td>
| + | |
- | </tr><tr>
| + | |
- | <td>BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)</td>
| + | |
- | <td>-</td><td>-</td><td>1</td><td>-</td>
| + | |
- | </tr><tr>
| + | |
- | <td>BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007)</td>
| + | |
- | <td>-</td><td>-</td><td>-</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>EcoRⅠ</td>
| + | |
- | <td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 4</td>
| + | |
- | <td>0.9</td><td>0.9</td><td>0.9</td><td>0.9</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>8</td><td>8</td><td>8</td><td>8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>10</td><td>10</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0824-1.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1~8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | :From left;
| + | |
- | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> ?
| + | |
- | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006) -> OK
| + | |
- | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007) -> OK
| + | |
- | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006) -> OK
| + | |
- | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> OK
| + | |
- | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006) -> OK
| + | |
- | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007) -> OK
| + | |
- | ::#BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006) -> None?
| + | |
- | |}
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0824-2.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>5~8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>9</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>2</td></tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>12</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | From the left;
| + | |
- | (single digestuion)
| + | |
- | ::5, BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> OK
| + | |
- | ::6, BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006) -> OK
| + | |
- | ::7, BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2007) -> 3400bp:Bad, 2200bp:OK
| + | |
- | ::8, BBa_J13002[http://partsregistry.org/Part:BBa_J13002](2006) -> OK
| + | |
- | |}
| + | |
- | | + | |
- | ===25 August, 2008===
| + | |
- | :(24/8)--->'''[[Team:Chiba/protocol/digestion|Digestion]]'''
| + | |
- | ::#Double Digestion:BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
| + | |
- | ::#Double Digesiton:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
| + | |
- | ::#Single Digestion:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
| + | |
- | ::#BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
| + | |
- | | + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1</td><td>2</td><td>3</td><td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>5</td><td>5</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>EcoRⅠ</td>
| + | |
- | <td>0.5</td><td>o.5</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>SpeⅠ</td>
| + | |
- | <td>0.5</td><td>0.5</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 1</td>
| + | |
- | <td>1</td><td>1</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 3</td>
| + | |
- | <td>-</td><td>-</td><td>1</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td>
| + | |
- | <td>1</td><td>1</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>2</td><td>2</td><td>7.5</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>10</td><td>10</td><td>10</td><td>1</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | :--->[[Team:Chiba/protocol/gelcheck|Gel Check]]
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0825-1.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1~3</td><td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>10</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>2</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>-</td><td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>12</td><td>6</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | :From right;
| + | |
- | ::#Double Digestion:BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) -> OK
| + | |
- | ::#Double Digesiton:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> ?(low)
| + | |
- | ::#Single Digestion:BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> ?(low)
| + | |
- | ::#BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007) -> OK(low)
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | --->[[Team:Chiba/protocol/digestion|Digestion]]
| + | |
- | (Double Digestion)
| + | |
- | ::#BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
| + | |
- | ::#BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
| + | |
- | ::#BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
| + | |
- | | + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1</td><td>2</td><td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>33ng/μl×60μl</td><td>33ng/μl×60μl</td><td>100ng/μl×20μl</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>PstⅠ</td>
| + | |
- | <td>2</td><td>2</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>XbaⅠ</td>
| + | |
- | <td>2</td><td>2</td><td>-</td>
| + | |
- | </tr>]
| + | |
- | <td>SpeⅠ</td>
| + | |
- | <td>-</td><td>-</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 2</td>
| + | |
- | <td>-</td><td>-</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 3</td>
| + | |
- | <td>10</td><td>10</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td>
| + | |
- | <td>10</td><td>10</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>16</td><td>16</td><td>16</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>100</td><td>100</td><td>52</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | ===26 August, 2008===
| + | |
- | :(8/25)--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0825-2.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1~3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>30</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>6</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>36</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | ::1, BBa_C0178[http://partsregistry.org/Part:BBa_C0178]
| + | |
- | :::Insert:609bp -> OK
| + | |
- | |}
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0825-3.JPG]]
| + | |
- | |
| + | |
- | ::2, BBa_C0170[http://partsregistry.org/Part:BBa_C0170]
| + | |
- | :::Insert:609bp -> OK
| + | |
- | |}
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0825-4.JPG]]
| + | |
- | |
| + | |
- | ::3, BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)
| + | |
- | ::: -> OK
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | --->'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]'''
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
| + | |
- | :::#5μl
| + | |
- | :::#5μl
| + | |
- | :::#27μl
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0825-5.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1~3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>2</td></tr>
| + | |
- | <tr>
| + | |
- | <td>H2O</td>
| + | |
- | <td>8</td></tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>12</td></tr>
| + | |
- | </table>
| + | |
- | | + | |
- | :From left;
| + | |
- | ::#insert:C0178 -> OK
| + | |
- | ::#insert:C0170 -> OK?
| + | |
- | ::#vector:J04500 -> OK
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/ligation/dephosphorylation|SAP]]'''
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>26</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>SAP</td>
| + | |
- | <td>1</td></tr>
| + | |
- | <tr>
| + | |
- | <td>SAP Buffer</td>
| + | |
- | <td>3</td></tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>30μL</td></tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
| + | |
- | :::Sample No.3->9μl
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0826.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No.3</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6</td></tr>
| + | |
- | </table>
| + | |
- | ::Sample No.3 -> OK ・・・30ng?
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/ligation/ligation|Ligation]]'''
| + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>(1)</td><td>(2)</td><td>(3)</td><td>(4)</td><td>(5)</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>insert①C0178</td>
| + | |
- | <td>1</td><td>-</td><td>-</td><td>1</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>insert②C0170</td>
| + | |
- | <td>-</td><td>1</td><td>-</td><td>-</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>vector③J04500</td>
| + | |
- | <td>-</td><td>-</td><td>2</td><td>2</td><td>2</td>
| + | |
- | </tr>]
| + | |
- | <td>ligase</td>
| + | |
- | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer</td>
| + | |
- | <td>2</td><td>2</td><td>2</td><td>2</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>6</td><td>6</td><td>5</td><td>4</td><td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| + | |
- | ::#insert①C0178
| + | |
- | ::#insert②C0170
| + | |
- | ::#vector③J04500
| + | |
- | ::#insert①C0178+vector③J04500
| + | |
- | ::#insert②C0170+vector③J04500
| + | |
- | | + | |
- | ===27 August, 2008===
| + | |
- | [[Team:Chiba/protocol/digestion|Digestion Test]]
| + | |
- | ::Sample DNA : BBa_R0010[http://partsregistry.org/Part:BBa_R0010]
| + | |
- | | + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1</td><td>2</td><td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>1</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>PstⅠ</td>
| + | |
- | <td>-</td><td>-</td><td>0.5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>EcoRⅠ</td>
| + | |
- | <td>-</td><td>0.5</td><td>0.5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 3</td>
| + | |
- | <td>-</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td>
| + | |
- | <td>-</td><td>-</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>-</td><td>7.5</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>1</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0827.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1</td><td>2</td><td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>1</td><td>5</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>1</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>H2O</td>
| + | |
- | <td>4</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6</td><td>6</td><td>6</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | :From left;<br>
| + | |
- | :#BBa_R0010
| + | |
- | :#Single Digestion
| + | |
- | :#Double Digestion
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :(26/8)--->'''[[Team:Chiba/protocol/PCR|Colony-PCR]]'''
| + | |
- | ::Colony PCR of 5 colonies from ligation plates (26/8-(4),(5)) and one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
| + | |
- | | + | |
- | :<table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>(4)-1~5</td><td>(5)-1~5</td><td>(F2620)</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>insert①C0178</td>
| + | |
- | <td>1</td><td>-</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>insert②C0170</td>
| + | |
- | <td>-</td><td>1</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>vector③J04500</td>
| + | |
- | <td>-</td><td>-</td><td>2</td>
| + | |
- | </tr>]
| + | |
- | <td>ligase</td>
| + | |
- | <td>1</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer</td>
| + | |
- | <td>2</td><td>2</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>6</td><td>6</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>10</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0827-2.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample DNA</td>
| + | |
- | <td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>12</td></tr>
| + | |
- | </table>
| + | |
- | :From left;<br>
| + | |
- | :1~5 insert:C0170 + vector:J04500 -> OK
| + | |
- | :6 F2620・・Positive Control -> OK
| + | |
- | :7~11 insert:C0178 + vector:J04500 -> OK
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | :--->(28/8)'''[[Team:Chiba/protocol/DNA Purification/sigma|Miniprep]]'''
| + | |
- | | + | |
- | | + | |
- | | + | |
- | '''[[Team:Chiba/protocol/transformation|Transformation]]'''
| + | |
- | :competent cells : XL10G
| + | |
- | ::*BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
| + | |
- | ::*BBa_C0062(2006) [http://partsregistry.org/Part:BBa_C0062] -> no colony on plates
| + | |
- | ::*BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
| + | |
- | ::*BBa_C0179(2006) [http://partsregistry.org/Part:BBa_C0179]
| + | |
- | ::*BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
| + | |
- | ::*BBa_S03156(2006) [http://partsregistry.org/Part:BBa_S03156]
| + | |
- | ::*BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
| + | |
- | ::*BBa_S03158(2006) [http://partsregistry.org/Part:BBa_S03158]
| + | |
- | ::*BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
| + | |
- | ::*BBa_S0316(2006) [http://partsregistry.org/Part:BBa_S03160]
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| + | |
- | | + | |
- | | + | |
- | :--->(29/8)'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | | + | |
- | | + | |
- | | + | |
- | ===28 August, 2008===
| + | |
- | :(27/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]
| + | |
- | ::#insert:C0170 + vector:J04500
| + | |
- | ::#insert:C0178 + vector:J04500
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/digestion|Digestion Test]]'''
| + | |
- | ::#insert:C0170 + vector:J04500
| + | |
- | ::#insert:C0178 + vector:J04500
| + | |
- | | + | |
- | <table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>3</td><td>4</td><td>5</td><td>6</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>insert:C0170 + vector:J04500 </td>
| + | |
- | <td>1</td><td>-</td><td>1</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>insert:C0178 + vector:J04500 </td>
| + | |
- | <td>-</td><td>1</td><td>-</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>EcoRⅠ</td>
| + | |
- | <td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>SpeⅠ</td>
| + | |
- | <td>-</td><td>-</td><td>0.1</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 3</td>
| + | |
- | <td>1</td><td>1</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td>
| + | |
- | <td>-</td><td>-</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>7.8</td><td>7.8</td><td>6.8</td><td>6.8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>10</td><td>10</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :--->(29/8)'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0828.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1,2</td><td>3~6</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>1</td><td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>1</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>H2O</td>
| + | |
- | <td>4</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6</td><td>12</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | :From left;
| + | |
- | ::# -> OK
| + | |
- | ::# -> OK
| + | |
- | ::# -> OK
| + | |
- | ::# -> OK
| + | |
- | ::# -> OK
| + | |
- | ::# -> OK
| + | |
- | |}
| + | |
- | | + | |
- | ===29 August, 2008===
| + | |
- | :--->'''[[Team:Chiba/protocol/digestion|Digestion Test]]'''
| + | |
- | ::#BBa_R0010(2007) [http://partsregistry.org/Part:BBa_R0010]
| + | |
- | ::#BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
| + | |
- | ::#BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
| + | |
- | ::#BBa_C0179(2006) [http://partsregistry.org/Part:BBa_C0179]
| + | |
- | ::#BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
| + | |
- | ::#BBa_S03156(2006) [http://partsregistry.org/Part:BBa_S03156]
| + | |
- | ::#BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
| + | |
- | ::#BBa_S03158(2006) [http://partsregistry.org/Part:BBa_S03158]
| + | |
- | ::#BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
| + | |
- | ::#BBa_S03160(2006) [http://partsregistry.org/Part:BBa_S03160]
| + | |
- | | + | |
- | | + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>S1~10</td><td>W1</td><td>W2~10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>1</td><td>4</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>PstⅠ</td>
| + | |
- | <td>-</td><td>0.075</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>EcoRⅠ</td>
| + | |
- | <td>0.1</td><td>0.075</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 3</td>
| + | |
- | <td>1</td><td>0.75</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td>
| + | |
- | <td>-</td><td>0.75</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>7.9</td><td>4.35</td><td>5.8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>10</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0829.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No.</td>
| + | |
- | <td>1~10</td><td>S1~S10,W1~W10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>1</td><td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>1</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6</td><td>12</td></tr>
| + | |
- | </table>
| + | |
- | :From left;<br>
| + | |
- | ::1~5,S1,S5,W1~W5
| + | |
- | |}
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0829-2.JPG]]
| + | |
- | |
| + | |
- | :From left;<br>
| + | |
- | ::6~10,S6~S10,W6~W10
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :(27/8)--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0829-3.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample DNA</td>
| + | |
- | <td>0.5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>4.5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6μL</td></tr>
| + | |
- | </table>
| + | |
- | | + | |
- | ::R0010 -> OK
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | '''[[Team:Chiba/protocol/digestion|Digestion]]'''
| + | |
- | ::#BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
| + | |
- | ::#BBa_I9030(2007) [http://partsregistry.org/Part:BBa_I9030]
| + | |
- | ::#BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
| + | |
- | ::#BBa_R0010(2006) [http://partsregistry.org/Part:BBa_R0010]
| + | |
- | | + | |
- | | + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1~3</td><td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>100ng/μL×20</td><td>200ng/μL×30</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>PstⅠ</td>
| + | |
- | <td>2</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>XbaⅠ</td>
| + | |
- | <td>2</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>SpeⅠ</td>
| + | |
- | <td>-</td><td>2</td></tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 2</td>
| + | |
- | <td>-</td><td>5</td></tr>
| + | |
- | <tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 3</td>
| + | |
- | <td>5</td><td>-</td></tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td>
| + | |
- | <td>5</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>16</td><td>6</td></tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>50</td><td>50</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0829-4.JPG]]
| + | |
- | |
| + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No.</td>
| + | |
- | <td>1~3</td><td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>10</td><td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>2</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>-</td><td>7</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>12</td><td>12</td></tr>
| + | |
- | </table>
| + | |
- | | + | |
- | :insert-I9026 : 685bp -> None<br>
| + | |
- | |}
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0829-5.JPG]]
| + | |
- | |
| + | |
- | :insert-I9030 : 745bp -> None<br>
| + | |
- | |}
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0829-6.JPG]]
| + | |
- | |
| + | |
- | :insert-S03154 : 685bp -> None<br>
| + | |
- | |}
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0829-7.JPG]]
| + | |
- | |
| + | |
- | :vector-R0010 : 2079bp -> OK<br>
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]'''
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
| + | |
- | :::->15μL
| + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/ligation/dephosphorylation|SAP]]'''
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>15</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>SAP</td>
| + | |
- | <td>2</td></tr>
| + | |
- | <tr>
| + | |
- | <td>SAP Buffer</td>
| + | |
- | <td>3</td></tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>10</td></tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>30μL</td></tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
| + | |
- | | + | |
- | ::--->Suspension! (We got information, by the team-output, to the effect that the density of the sample is too low.)
| + | |
- | | + | |
- | | + | |
- | ===30 August, 2008===
| + | |
- | :'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| + | |
- | :competent cells : XL10G
| + | |
- | ::*BBa_I9026(2007) [http://partsregistry.org/Part:BBa_I9026]
| + | |
- | ::*BBa_I9030(2006) [http://partsregistry.org/Part:BBa_I9030]
| + | |
- | ::*BBa_S03154(2007) [http://partsregistry.org/Part:BBa_S03154]
| + | |
- | ::*BBa_S03156(2007) [http://partsregistry.org/Part:BBa_S03156]
| + | |
- | ::*BBa_S03158(2007) [http://partsregistry.org/Part:BBa_S03158]
| + | |
- | ::*BBa_S03160(2007) [http://partsregistry.org/Part:BBa_S03160]
| + | |
- | ::*BBa_C0062(2007) [http://partsregistry.org/Part:BBa_C0062]
| + | |
- | ::*BBa_C0179(2007) [http://partsregistry.org/Part:BBa_C0179]
| + | |
- | | + | |
- | :--->(31/8)'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :(29/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| + | |
- | ::#insert:C0170 + vector:J04500
| + | |
- | ::#insert:C0178 + vector:J04500
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/digestion|Digestion Test]]'''
| + | |
- | ::*insert:C0170 + vector:J04500 -> Sample No.1~4
| + | |
- | ::*insert:C0178 + vector:J04500 -> Sample No.5~8
| + | |
- | ::*BBa_T9002(2007)[http://partsregistry.org/Part:BBa_T9002] -> Sample No.9
| + | |
- | ::*Single Digestion of Sample No.1~9 -> Sample No.10~18
| + | |
- | ::*Double Digestion of Sample No.1~9 -> Sample No.19~27
| + | |
- | | + | |
- | :<table width="500" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="270">Sample No.</td><td>Single : 10~18</td><td>Double : 19~27</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td><td>1</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>XbaⅠ</td><td>0.1</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>SpeⅠ</td><td>-</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 2</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td><td>6.9</td><td>2.8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0830.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1~9</td><td>10~18</td><td>19~27</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>1</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>1</td><td>2</td><td>2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>4</td><td>-</td><td>-</td></tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6</td><td>12</td><td>12</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | :From left;<br>
| + | |
- | ::Sanple No.1~13
| + | |
- | ::-> OK
| + | |
- | |}
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0830-2.JPG]]
| + | |
- | |
| + | |
- | :From left;<br>
| + | |
- | ::Sample No.14~17,24~16
| + | |
- | ::14 -> OK
| + | |
- | ::15,16 -> Bad
| + | |
- | ::17 -> None
| + | |
- | ::24~16 -> Bad
| + | |
- | |}
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0830-3.JPG]]
| + | |
- | |
| + | |
- | :From left;<br>
| + | |
- | ::Sample No.18,27,19~23
| + | |
- | ::18,27 -> Bad
| + | |
- | ::19~22 -> OK
| + | |
- | ::23 -> OK??
| + | |
- | |}
| + | |
- | | + | |
- | ==Week 3==
| + | |
- | ===31 August, 2008===
| + | |
- | :'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| + | |
- | :Competent Cells : XL10G
| + | |
- | ::*Plac+RBS+LasI
| + | |
- | ::*Plac+RBS+RhlI
| + | |
- | | + | |
- | '''[[Team:Chiba/protocol/digestion|Digestion]]'''
| + | |
- | ::#BBa_I9026[http://partsregistry.org/Part:BBa_I9026](2007)
| + | |
- | ::#BBa_I9030[http://partsregistry.org/Part:BBa_I9030](2006)
| + | |
- | ::#BBa_S03154[http://partsregistry.org/Part:BBa_S03154](2007)
| + | |
- | ::#BBa_R0010[http://partsregistry.org/Part:BBa_R0010](2007)
| + | |
- | | + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1~3</td><td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td><td>12</td><td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>PstⅠ</td>
| + | |
- | <td>0.2</td><td>0.2</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>XbaⅠ</td><td>0.2</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>SpeⅠ</td><td>-</td><td>0.4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 2</td>
| + | |
- | <td>-</td><td>1.5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 3</td>
| + | |
- | <td>2</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td>
| + | |
- | <td>2</td><td>1.5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>3.6</td><td>1.4</td></tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>20</td><td>15</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | :--->(1/9)'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | | + | |
- | | + | |
- | :(30/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| + | |
- | | + | |
- | | + | |
- | :--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
| + | |
- | :*Plac+RBS+RhlI Sample No.2~5 | + | |
- | :*BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、② | + | |
- | | + | |
- | <table width="600" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample</td>
| + | |
- | <td>Single Digesiton 2~5</td><td>Double Digestion 2~5</td><td>Single Digestion T9002①</td><td>Single Digesiton T9002②</td><td><Double Digestion T9002①②</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td><td>1</td><td>3</td><td>3</td><td>1</td><td>3</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>XbaⅠ</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>SpeⅠ</td><td>-</td><td>0.1</td><td>-</td><td>-</td><td>0.1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Buffer 2</td>
| + | |
- | <td>0.9</td><td>0.8</td><td>0.9</td><td>0.9</td><td>0.8</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>BSA</td>
| + | |
- | <td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>7</td><td>5</td><td>5</td><td>7</td><td>5</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | | + | |
- | | + | |
- | --->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | | + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0831-p87.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>Plac+RBS+RhlI Sample No.2~5, BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①②
| + | |
- | </td><td>Single & Double Digestion</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>1</td><td>10</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>1</td><td>2</td></tr>
| + | |
- | <tr>
| + | |
- | <td>dH2O</td>
| + | |
- | <td>4</td><td>-</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>6</td><td>12</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | :From Left
| + | |
- | ::*Single Digestion Plac+RBS+RhlI 2~5
| + | |
- | ::*Single Digestion BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、②
| + | |
- | ::*Double Digestion Plac+RBS+RhlI 2~5
| + | |
- | ::*Double Digestion BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、②
| + | |
- | ::*BBa_T9002[http://partsregistry.org/Part:BBa_T9002]①、②
| + | |
- | |}
| + | |
- | | + | |
- | ===1 September, 2008===
| + | |
- | :(31/8)--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0901.JPG]]
| + | |
- | |
| + | |
- | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
| + | |
- | <tr>
| + | |
- | <td width="257">Sample No</td>
| + | |
- | <td>1~3</td><td>4</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Sample DNA</td>
| + | |
- | <td>20</td><td>15</td>
| + | |
- | </tr>
| + | |
- | <tr>
| + | |
- | <td>Loading Dye</td>
| + | |
- | <td>4</td><td>3</td></tr>
| + | |
- | <tr>
| + | |
- | <td>TOTAL</td>
| + | |
- | <td>24</td><td>18</td>
| + | |
- | </tr>
| + | |
- | </table>
| + | |
- | :From left;
| + | |
- | ::insert-1(I9026)
| + | |
- | ::insert-2(I9030)
| + | |
- | |}
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0901-2.JPG]]
| + | |
- | |
| + | |
- | ::insert-3(S03154)
| + | |
- | |}
| + | |
- | {|align="justify"
| + | |
- | |[[Image:Chiba-0901-3.JPG]]
| + | |
- | |
| + | |
- | ::vector-4(R0010)
| + | |
- | |}
| + | |
- | | + | |
- | | + | |
- | ===2 September, 2008===
| + | |
- | | + | |
- | ===3 September, 2008===
| + | |
- | | + | |
- | ===4 September, 2008===
| + | |
- | | + | |
- | ===5 September, 2008===
| + | |
- | | + | |
- | ===6 September, 2008===
| + | |
- | | + | |
- | ==Week 4==
| + | |
- | ===7 September, 2008===
| + | |
- | | + | |
- | ===8 September, 2008===
| + | |
- | | + | |
- | ===9 September, 2008===
| + | |
- | | + | |
- | ===10 September, 2008===
| + | |
- | | + | |
- | ===11 September, 2008===
| + | |
- | | + | |
- | ===12 September, 2008===
| + | |
- | | + | |
- | ===13 September, 2008===
| + | |
- | | + | |
- | ==Week 5==
| + | |
- | ===14 September, 2008===
| + | |
- | :(13/9)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
| + | |
- | | + | |
| | | |
| | | |
- | :'''[[Team:Chiba/protocol/transformation|Transformation]]'''
| + | ===Cross-Talk Check=== |
- | :Competent cells : XL10G
| + | #LuxR vs LuxI('''OK'''),LasI('''OK'''),CinI('''No'''),RhlI('''OK''') |
- | ::*BBa_T9002[http://partsregistry.org/Part:BBa_T9002](2007)
| + | #LuxI vs LuxR,LasR,CinR,RhlR-->発現を確認できず |
- | ::*Plac+RhlI+LVA
| + | |
- | ::*Plac+CinI+LVA
| + | |
| | | |
- | :--->(15/9)
| + | ===time-delay test=== |
| + | [[Team:Chiba/protocol/phenotype/timedelay/j|time-delay protocol]] |
| | | |
- | ===15 September, 2008=== | + | ==連絡== |
| + | ===pptに載せたいもの=== |
| + | ====作ったもの==== |
| + | *絵で、わかりやすく。~Isごとに色を変える。 |
| + | ====time-delay実験結果の整理(25/9)==== |
| + | *実験操作を分かりやすく書く |
| + | *比較するもの |
| + | **LuxI,LasI,RhlI |
| + | **TAG有/無 |
| + | **温度 |
| + | **菌数比 |
| + | **株 |
| + | *ポイント |
| + | #正しく書きだす |
| + | #推論を立てる(帰納的に、結論を急がないこと) |
| + | ====固体培地spot実験(28/9)==== |
| + | 前日 |
| + | *IPTG入りLB-Agar-amp(20ml)に、T9002を2ml(10<sup>3</sup>-10<sup>5</sup> cells/2ml)撒く-->37℃,O/N |
| + | *~Is(5種類)をLB-Amp,2ml培養 |
| + | 当日 |
| + | *T9002がコロニーを形成したプレートの中心に、~Isの培養液を1μl,spotする。 |
| + | *gfpの発現が同心円状に広がっていく。その計時変化を記録する。 |
| | | |
- | ===16 September, 2008===
| |
| | | |
- | {| style="color:white;background-color:Maroon" cellpadding="3" cellspacing="3" border="1" bordercolor="white" width="100%" align="center" | + | {| style="color:white;" cellpadding="3" cellspacing="3" border="0" width="100%" align="center" class="menu" | |
| !align="center"|[[Team:Chiba/j|ホーム]] | | !align="center"|[[Team:Chiba/j|ホーム]] |
| !align="center"|[[Team:Chiba/Team/j|メンバー紹介]] | | !align="center"|[[Team:Chiba/Team/j|メンバー紹介]] |
| !align="center"|[[Team:Chiba/Project/j|プロジェクト紹介]] | | !align="center"|[[Team:Chiba/Project/j|プロジェクト紹介]] |
- | !align="center"|[[Team:Chiba/Parts/j|Parts Submitted to the Registry]] | + | !align="center"|[[Team:Chiba/Parts/j|作成したDNAパーツ]] |
- | !align="center"|[[Team:Chiba/Modeling/j|モデリング]]
| + | |
| !align="center"|[[Team:Chiba/Internal|ノート]] | | !align="center"|[[Team:Chiba/Internal|ノート]] |
| |} | | |} |
任意の順番と、delay-timeをもった遺伝子発現を実現するために、バクテリア間の細胞間通信、クオラムセンシングを利用する。クオラムセンシングでは、応答閾値を超えるAHLが蓄積されたとき初めて、目的遺伝子の発現がおこる。AHL情報がじゅうぶんゆっくり蓄積するとき、ひとつのAHL送信装置に対して、感度の異なる複数の受信装置によってさまざまな出力装置を独立に起動すれば、感度の高いAHL 受信機ほど先に起動する。