Team:Chiba/jk/β/week 2

>[[Team:Chiba/jk/β|送受信班]]

==Week 2==
===[[Team:Chiba/jk/β/week 1|>last week]]===
===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="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>5</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>0.5</td>
</tr>
<tr>
<td>Thermopol Buffer</td>
<td>5</td>
</tr>
<tr>
<td>dH2O</td>
<td>28.5</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-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??
|}

===[[Team:Chiba/jk/β/week 3|>next week]]===

{| style="color:white;background-color:Maroon" cellpadding="3" cellspacing="3" border="1" bordercolor="white" width="100%" align="center"
!align="center"|[[Team:Chiba/j|ホーム]]
!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/Parts/j|Parts Submitted to the Registry]]
!align="center"|[[Team:Chiba/Modeling/j|モデリング]]
!align="center"|[[Team:Chiba/Internal|ノート]]
|}