Team:Chiba/jk/β
From 2008.igem.org
(Difference between revisions)
(→23 August, 2008) |
(→Week 2) |
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==Week 2== | ==Week 2== | ||
- | + | ===24 August, 2008=== | |
- | :(22/8)--->[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]] | + | :(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"> | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
<td width="257">Sample No</td> | <td width="257">Sample No</td> | ||
- | <td> | + | <td>5</td><td>6</td><td>7</td><td>8</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007)</td> |
- | + | <td>1</td><td>-</td><td>-</td><td>-</td> | |
</tr> | </tr> | ||
<tr> | <tr> | ||
- | <td> | + | <td>BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2006)</td> |
- | <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> | ||
<tr> | <tr> | ||
<td>dH2O</td> | <td>dH2O</td> | ||
- | <td> | + | <td>8</td><td>8</td><td>8</td><td>8</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>10</td><td>10</td><td>10</td><td>10</td> |
</tr> | </tr> | ||
</table> | </table> | ||
+ | |||
+ | |||
+ | |||
+ | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | ||
{|align="justify" | {|align="justify" | ||
|[[Image:Chiba-0824-1.JPG]] | |[[Image:Chiba-0824-1.JPG]] | ||
| | | | ||
- | + | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | |
- | : | + | |
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | + | ||
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | + | |
<tr> | <tr> | ||
<td width="257">Sample No</td> | <td width="257">Sample No</td> | ||
- | <td> | + | <td>1~8</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Sample DNA</td> | <td>Sample DNA</td> | ||
- | <td> | + | <td>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Loading Dye</td> | <td>Loading Dye</td> | ||
- | <td> | + | <td>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>dH2O</td> | <td>dH2O</td> | ||
- | <td> | + | <td>4</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td> | + | <td>6</td> |
</tr> | </tr> | ||
</table> | </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 | ||
+ | (↓single digestuion) | ||
+ | ::#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" | {|align="justify" | ||
|[[Image:Chiba-0824-2.JPG]] | |[[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; | From the left; | ||
(single digestuion) | (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]] | :(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"> | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
<td width="257">Sample No</td> | <td width="257">Sample No</td> | ||
- | <td> | + | <td>1</td><td>2</td><td>3</td><td>4</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 882: | Line 925: | ||
:--->[[Team:Chiba/protocol/gelcheck|Gel Check]] | :--->[[Team:Chiba/protocol/gelcheck|Gel Check]] | ||
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | + | {|align="justify" |
+ | |[[Image:Chiba-0825-1.JPG]] | ||
+ | | | ||
+ | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
<td width="257">Sample No</td> | <td width="257">Sample No</td> | ||
- | <td> | + | <td>1~3</td><td>4</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Sample DNA</td> | <td>Sample DNA</td> | ||
- | <td>10</td><td> | + | <td>10</td><td>1</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Loading Dye</td> | <td>Loading Dye</td> | ||
- | + | <td>2</td><td>1</td> | |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>dH2O</td> | <td>dH2O</td> | ||
- | + | <td>-</td><td>4</td> | |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | + | <td>12</td><td>6</td> | |
</tr> | </tr> | ||
</table> | </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) | |
- | From right; | + | ::#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]] | --->[[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"> | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
<td width="257">Sample No</td> | <td width="257">Sample No</td> | ||
- | <td> | + | <td>1</td><td>2</td><td>3</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
Line 962: | Line 1,006: | ||
</table> | </table> | ||
- | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | + | |
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | + | |
+ | ===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> | <tr> | ||
<td width="257">Sample No</td> | <td width="257">Sample No</td> | ||
- | <td> | + | <td>1~3</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Sample DNA</td> | <td>Sample DNA</td> | ||
- | + | <td>30</td> | |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Loading Dye</td> | <td>Loading Dye</td> | ||
- | + | <td>6</td> | |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | + | <td>36</td> | |
</tr> | </tr> | ||
</table> | </table> | ||
- | + | ::1, BBa_C0178[http://partsregistry.org/Part:BBa_C0178] | |
- | + | ||
- | + | ||
- | : | + | |
:::Insert:609bp -> OK | :::Insert:609bp -> OK | ||
|} | |} | ||
Line 990: | Line 1,037: | ||
|[[Image:Chiba-0825-3.JPG]] | |[[Image:Chiba-0825-3.JPG]] | ||
| | | | ||
- | :: | + | ::2, BBa_C0170[http://partsregistry.org/Part:BBa_C0170] |
:::Insert:609bp -> OK | :::Insert:609bp -> OK | ||
|} | |} | ||
Line 996: | Line 1,043: | ||
|[[Image:Chiba-0825-4.JPG]] | |[[Image:Chiba-0825-4.JPG]] | ||
| | | | ||
- | :: | + | ::3, BBa_J04500[http://partsregistry.org/Part:BBa_J04500](2007) |
::: -> OK | ::: -> OK | ||
|} | |} | ||
- | + | --->'''[[Team:Chiba/protocol/ligation/gelex|Gel Extract]]''' | |
- | + | ||
:--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]''' | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]''' | ||
- | ::: | + | :::#5μl |
- | ::: | + | :::#5μl |
- | ::: | + | :::#27μl |
+ | |||
:--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | ||
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | + | {|align="justify" |
+ | |[[Image:Chiba-0825-5.JPG]] | ||
+ | | | ||
+ | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
<td width="257">Sample No</td> | <td width="257">Sample No</td> | ||
- | <td> | + | <td>1~3</td> |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Sample DNA</td> | <td>Sample DNA</td> | ||
- | + | <td>2</td> | |
</tr> | </tr> | ||
<tr> | <tr> | ||
<td>Loading Dye</td> | <td>Loading Dye</td> | ||
- | <td>2</td> | + | <td>2</td></tr> |
- | </tr> | + | |
<tr> | <tr> | ||
<td>H2O</td> | <td>H2O</td> | ||
- | <td>8</td> | + | <td>8</td></tr> |
- | </tr> | + | |
<tr> | <tr> | ||
<td>TOTAL</td> | <td>TOTAL</td> | ||
- | <td>12</td> | + | <td>12</td></tr> |
- | </tr> | + | |
</table> | </table> | ||
- | + | :From left; | |
- | + | ::#insert:C0178 -> OK | |
- | + | ::#insert:C0170 -> OK? | |
- | From left; | + | ::#vector:J04500 -> OK |
- | :: | + | |
- | :: | + | |
- | :: | + | |
|} | |} | ||
- | :--->''' | + | :--->'''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]]''' | :--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]''' | ||
- | ::: | + | :::Sample No.3->9μl |
:--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | :--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]''' | ||
- | <table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | + | {|align="justify" |
+ | |[[Image:Chiba-0826.JPG]] | ||
+ | | | ||
+ | :<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000"> | ||
<tr> | <tr> | ||
- | <td width="257">Sample | + | <td width="257">Sample No.3</td> |
<td>1</td> | <td>1</td> | ||
</tr> | </tr> | ||
Line 1,069: | Line 1,136: | ||
<td>6</td></tr> | <td>6</td></tr> | ||
</table> | </table> | ||
- | + | ::Sample No.3 -> OK ・・・30ng? | |
- | + | ||
- | + | ||
- | + | ||
|} | |} | ||
Line 1,113: | Line 1,177: | ||
:--->'''[[Team:Chiba/protocol/transformation|Transformation]]''' | :--->'''[[Team:Chiba/protocol/transformation|Transformation]]''' | ||
- | :: | + | ::#insert①C0178 |
- | :: | + | ::#insert②C0170 |
- | :: | + | ::#vector③J04500 |
- | :: | + | ::#insert①C0178+vector③J04500 |
- | :: | + | ::#insert②C0170+vector③J04500 |
- | + | ===27 August, 2008=== | |
[[Team:Chiba/protocol/digestion|Digestion Test]] | [[Team:Chiba/protocol/digestion|Digestion Test]] | ||
::Sample DNA : BBa_R0010[http://partsregistry.org/Part:BBa_R0010] | ::Sample DNA : BBa_R0010[http://partsregistry.org/Part:BBa_R0010] | ||
Line 1,128: | Line 1,192: | ||
<tr> | <tr> | ||
<td width="257">Sample No</td> | <td width="257">Sample No</td> | ||
- | <td> | + | <td>1</td><td>2</td><td>3</td> |
</tr> | </tr> | ||
<tr> | <tr> |
Revision as of 16:30, 11 September 2008
β班の実験ノート
Contents |
Week 1
17 August, 2008
18 August, 2008
DNA Template | 1 |
dNTP mix | 10 |
Foward Primer | 5 |
Reverse Primer | 5 |
DNA polymerase | 1 |
Thermopol Buffer | 10 |
dH2O | 68 |
TOTAL | 100μL |
- 95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃
- --->Gel Check
compitent cell : XL10G
- --->(20/8)Mini prep
19 August, 2008
DNA Template | 1 |
dNTP mix | 10 |
Foward Primer | 5 |
Reverse Primer | 5 |
DNA polymerase VENT | 1 |
Thermopol Buffer | 5 |
dH2O | 28 |
TOTAL | 50μL |
- 95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃
- --->Gel Check
20 August, 2008
- (18/8)--->Mini prep
- --->Gel Check
--->[Team:Chiba/protocol/digestion|Digestion test]]
Sample No | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 |
BBa_C0170(2006) [68] | 3 | - | - | - | 3 | - | - | - |
BBa_C0170(2007) [69] | - | 3 | - | - | - | 3 | - | - |
BBa_C0178(2006) [70] | - | - | 3 | - | - | - | 3 | - |
BBa_C0178(2007) [71] | - | - | - | 3 | - | - | - | 3 |
EcoRⅠ | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 | 0.1 |
PstⅠ | - | - | - | - | 0.1 | 0.1 | 0.1 | 0.1 |
Buffer EcoRⅠ | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
BSA | 1 | 1 | 1 | 1 | 1 | 1 | 1 | 1 |
dH2O | 4.9 | 4.9 | 4.9 | 4.9 | 4.8 | 4.8 | 4.8 | 4.8 |
TOTAL | 10 | 10 | 10 | 10 | 10 | 10 | 10 | 10 |
- --->Gel Check
|
21 August, 2008
- (20/8)--->PCR
DNA Template | 1 |
dNTP mix | 10 |
Foward Primer | 5 |
Reverse Primer | 5 |
DNA polymerase TAQ | 1 |
Thermopol Buffer | 5 |
dH2O | 28 |
TOTAL | 50μL |
- 95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )・・・25cycles -> 72℃,10min -> 6℃
- --->Gel Check
Compitentcell : XL10G
--->(23/8)Digestion
22 August, 2008
- Transformation
- (Locate & 2 Punch ~iyama ver.~)
- Compitentcell : XL10G
--->(24/8)Mini prep
23 August, 2008
- (21/8)--->Mini prep
- --->Digestion test
Sample No 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 BBa_I9030(2007) [117] 3 - - - - - - 3 - - - - - - 3 - - - - - - BBa_I9030(2006) [118] - 3 - - - - - - 3 - - - - - - 3 - - - - - BBa_S03154(2007) [119] - - 3 - - - - - - 3 - - - - - - 3 - - - - BBa_S03154(2006) [120] - - - 3 - - - - - - 3 - - - - - - 3 - - - BBa_I9026(2007) [121] - - - - 3 - - - - - - 3 - - - - - - 3 - - BBa_I9026(2007) [122] - - - - - 3 - - - - - - 3 - - - - - - 3 - BBa_J04500(2007) [123] - - - - - - 1 - - - - - - 3 - - - - - - 3 EcoRⅠ - - - - - - - 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 0.1 PstⅠ - - - - - - - - - - - - - - 0.1 0.1 0.1 0.1 0.1 0.1 0.1 Buffer EcoRⅠ - - - - - - - 0.9 0.9 0.9 0.9 0.9 0.9 0.9 0.8 0.8 0.8 0.8 0.8 0.8 0.8 BSA - - - - - - - 1 1 1 1 1 1 1 1 1 1 1 1 1 1 dH2O - - - - - - - 5 5 5 5 5 5 5 5 5 5 5 5 5 5 TOTAL 3 3 3 3 3 3 1 10 10 10 10 10 10 10 10 10 10 10 10 10 10
- --->Gel Check
Sample No 8~21 Sample DNA 10 Loading Dye 2 TOTAL 12 - From left;
- (Single Digestion) Sanple No. 8~14
- Digestion test
- BBa_J04500(2007:old-sample)[145]
Sample No 1 2 3 Sample DNA 3 3 3 EcoRⅠ - 0.1 0.1 SpeⅠ - - 0.1 EcoRⅠ Buffer - 0.9 0.8 BSA - 1 1 dH2O - 5 5 TOTAL 3 10 10
--->Gel CheckWeek 2
24 August, 2008
- (22/8)--->Mini prep
- --->Digestion test
Sample No 5 6 7 8 BBa_J04500[157](2007) 1 - - - BBa_J04500[158](2006) - 1 - - BBa_J13002[159](2007) - - 1 - BBa_J13002[160](2007) - - - 1 EcoRⅠ 0.1 0.1 0.1 0.1 Buffer 4 0.9 0.9 0.9 0.9 dH2O 8 8 8 8 TOTAL 10 10 10 10
- --->Gel Check
Sample No 1~8 Sample DNA 1 Loading Dye 1 dH2O 4 TOTAL 6 - From left;
(↓single digestuion)
25 August, 2008
- (24/8)--->Digestion
Sample No 1 2 3 4 Sample DNA 5 5 1 1 EcoRⅠ 0.5 o.5 - - SpeⅠ 0.5 0.5 - - Buffer 1 1 1 - - Buffer 3 - - 1 - BSA 1 1 - - dH2O 2 2 7.5 - TOTAL 10 10 10 1 - --->Gel Check
--->Digestion (Double Digestion)Sample No 1 2 3 Sample DNA 33ng/μl×60μl 33ng/μl×60μl 100ng/μl×20μl PstⅠ 2 2 2 XbaⅠ 2 2 - SpeⅠ - - 2 </tr>
Buffer 2 - - 5 Buffer 3 10 10 - BSA 10 10 5 dH2O 16 16 16 TOTAL 100 100 52
26 August, 2008
- (8/25)--->Gel Check
Sample No 1~3 Sample DNA 30 Loading Dye 6 TOTAL 36 - 1, BBa_C0178[184]
- Insert:609bp -> OK
- 1, BBa_C0178[184]
- 2, BBa_C0170[185]
- Insert:609bp -> OK
- 2, BBa_C0170[185]
- 3, BBa_J04500[186](2007)
- -> OK
- 3, BBa_J04500[186](2007)
--->Gel Extract
- --->Zymo Clean
- 5μl
- 5μl
- 27μl
- --->Gel Check
Sample No 1~3 Sample DNA 2 Loading Dye 2 H2O 8 TOTAL 12 - From left;
- insert:C0178 -> OK
- insert:C0170 -> OK?
- vector:J04500 -> OK
- From left;
- --->SAP
Sample No 3 Sample DNA 26 SAP 1 SAP Buffer 3 TOTAL 30μL
- --->Zymo Clean
- Sample No.3->9μl
- --->Gel Check
Sample No.3 1 Loading Dye 1 dH2O 4 TOTAL 6 - Sample No.3 -> OK ・・・30ng?
- --->Ligation
Sample No (1) (2) (3) (4) (5) insert①C0178 1 - - 1 - insert②C0170 - 1 - - 1 vector③J04500 - - 2 2 2 ligase 1 1 1 1 1 </tr>
Buffer 2 2 2 2 2 dH2O 6 6 5 4 4 TOTAL 10 10 10 10 10
- --->Transformation
- insert①C0178
- insert②C0170
- vector③J04500
- insert①C0178+vector③J04500
- insert②C0170+vector③J04500
27 August, 2008
- Sample DNA : BBa_R0010[187]
Sample No 1 2 3 Sample DNA 1 1 1 PstⅠ - - 0.5 EcoRⅠ - 0.5 0.5 Buffer 3 - 1 1 BSA - - 1 dH2O - 7.5 2 TOTAL 1 10 10
- --->Gel Check
Sample No ① ② ③ Sample DNA 1 5 5 Loading Dye 1 1 1 H2O 4 - - TOTAL 6 6 6 From left;
- ①BBa_R0010
- ②Single Digestion
- ③Double Digestion
- (26/8)--->PCR
- Colony PCR of 5 colonies from ligation plates (26/8-(4),(5)) and one from control plate(BBa_F2620[188](2007)).
Sample No (4)-1~5 (5)-1~5 (F2620) insert①C0178 1 - - insert②C0170 - 1 - vector③J04500 - - 2 ligase 1 1 1 </tr>
Buffer 2 2 2 dH2O 6 6 5 TOTAL 10 10 10 - --->Gel Check
Sample DNA 10 Loading Dye 2 TOTAL 12 From left;
- 1~5 insert:C0170 + vector:J04500 -> OK
- 6 F2620・・Positive Control -> OK
- 7~11 insert:C0178 + vector:J04500 -> OK
- --->(28/8)Mini prepDigestion Test
Transformation comptent cell : XL10G- --->Mini prep
- --->(29/8)Gel Check
- 28 August, 2008]
(27/8)--->Mini prep
- ①insert:C0170 + vector:J04500
- ②insert:C0178 + vector:J04500
- --->Digestion Test
- --->(29/8)Gel Check
Sample No ①② Single Digesiton:①② Double Digesiton:①② Sample DNA 1 10 10 Loading Dye 1 2 2 H2O 4 - - TOTAL 6 12 12 From left;
- ① -> OK
- ② -> OK
- Single Digestion:① -> OK
- Single Digestion:② -> OK
- Double Digestion:① -> OK
- Double Digestion:② -> OK
- 29 August, 2008
- --->Digestion Test
Sample No S①~⑩ w① w②~⑩ Sample DNA 1 4 2 PstⅠ - 0.075 0.1 EcoRⅠ 0.1 0.075 0.1 Buffer 3 1 0.75 1 BSA - 0.75 1 dH2O 7.9 4.35 5.8 TOTAL 10 10 10 - --->Gel Check
Sample DNA 10 Loading Dye 2 TOTAL 12 From left;
①~⑤,S①⑤,W①~⑤From left;
⑥~⑩,S⑥~⑩,W⑥~⑩
(27/8)--->R0010
Sample No S①~③ ④ Sample DNA 100ng/μL×20 200ng/μL×30 PstⅠ 2 2 XbaⅠ 2 - SpeⅠ - 2 Buffer 2 - 5 Buffer 3 5 - BSA 5 5 dH2O 16 6 TOTAL 50 50 - --->Gel Check
Sample DNA 10 3 Loading Dye 2 2 dH2O - 7 TOTAL 12
From left;
From left;
From left;
From left;
)--->Gel Extract- --->Zymo Clean
- =>1.5μL/2colum
- --->SAP
④
- --->Zymo Clean
- ---> miss!
- 30 August, 2008
Transformation comptent cell : XL10G
- --->(31/8)Mini prep
(29/8)--->Mini prep- ①insert:C0170 + vector:J04500
- ②insert:C0178 + vector:J04500
- --->Digestion Test
- --->Gel Check
From left;
From left;
From left;
Week 3
- 31 August, 2008
- 1 September, 2008
- 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
ホーム メンバー紹介 プロジェクト紹介 Parts Submitted to the Registry モデリング ノート - --->Zymo Clean