Team:Chiba/jk/β/weak 2

From 2008.igem.org

(Difference between revisions)
Line 1: Line 1:
>[[Team:Chiba/jk/β|送受信班]]
>[[Team:Chiba/jk/β|送受信班]]
 +
==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="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??
 +
|}
{| style="color:white;background-color:Maroon" cellpadding="3" cellspacing="3" border="1" bordercolor="white" width="100%" align="center"
{| style="color:white;background-color:Maroon" cellpadding="3" cellspacing="3" border="1" bordercolor="white" width="100%" align="center"

Revision as of 07:17, 20 September 2008

>送受信班

Contents

Week 2

24 August, 2008

(22/8)--->Mini prep
  1. BBa_J04500[1](2007)
  2. BBa_J04500[2](2006)
  3. BBa_J13002[3](2007)
  4. BBa_J13002[4](2006)


--->Digestion test
  1. BBa_J04500[5](2007)
  2. BBa_J04500[6](2006)
  3. BBa_J13002[7](2007)
  4. BBa_J13002[8](2006)


Sample No 5678
BBa_J04500[9](2007) 1---
BBa_J04500[10](2006) -1--
BBa_J13002[11](2007) --1-
BBa_J13002[12](2007) ---1
EcoRⅠ 0.10.10.10.1
Buffer 4 0.90.90.90.9
dH2O 8888
TOTAL 10101010


--->Gel Check
Chiba-0824-1.JPG
Sample No 1~8
Sample DNA 1
Loading Dye 1
dH2O 4
TOTAL 6
From left;
  1. BBa_J04500[13](2007) -> ?
  2. BBa_J04500[14](2006) -> OK
  3. BBa_J13002[15](2007) -> OK
  4. BBa_J13002[16](2006) -> OK
  5. BBa_J04500[17](2007) -> OK
  6. BBa_J04500[18](2006) -> OK
  7. BBa_J13002[19](2007) -> OK
  8. BBa_J13002[20](2006) -> None?
Chiba-0824-2.JPG
Sample No 5~8
Sample DNA 9
Loading Dye 2
dH2O 1
TOTAL 12

From the left; （single digestuion）

5, BBa_J04500[21](2007) -> OK
6, BBa_J04500[22](2006) -> OK
7, BBa_J13002[23](2007) -> 3400bp:Bad, 2200bp:OK
8, BBa_J13002[24](2006) -> OK

25 August, 2008

(24/8)--->Digestion
  1. Double Digestion:BBa_J04500[25](2007)
  2. Double Digesiton:BBa_R0010[26](2007)
  3. Single Digestion:BBa_R0010[27](2007)
  4. BBa_R0010[28](2007)
Sample No 1234
Sample DNA 5511
EcoRⅠ 0.5o.5--
SpeⅠ 0.50.5--
Buffer 1 11--
Buffer 3 --1-
BSA 11--
dH2O 227.5-
TOTAL 1010101
--->Gel Check
Chiba-0825-1.JPG
Sample No 1~34
Sample DNA 101
Loading Dye 21
dH2O -4
TOTAL 126
From right;
  1. Double Digestion:BBa_J04500[29](2007) -> OK
  2. Double Digesiton:BBa_R0010[30](2007) -> ?(low)
  3. Single Digestion:BBa_R0010[31](2007) -> ?(low)
  4. BBa_R0010[32](2007) -> OK(low)


--->Digestion (Double Digestion)

  1. BBa_C0178[33]
  2. BBa_C0170[34]
  3. BBa_J04500[35](2007)
]

</tr>

Sample No 123
Sample DNA 33ng/μl×60μl33ng/μl×60μl100ng/μl×20μl
PstⅠ 222
XbaⅠ 22-
SpeⅠ --2
Buffer 2 --5
Buffer 3 1010-
BSA 10105
dH2O 161616
TOTAL 10010052

26 August, 2008

(8/25)--->Gel Check
Chiba-0825-2.JPG
Sample No 1~3
Sample DNA 30
Loading Dye 6
TOTAL 36
1, BBa_C0178[36]
Insert:609bp -> OK
Chiba-0825-3.JPG
2, BBa_C0170[37]
Insert:609bp -> OK
Chiba-0825-4.JPG
3, BBa_J04500[38](2007)
-> OK


--->Gel Extract


--->Zymo Clean
  1. 5μl
  2. 5μl
  3. 27μl


--->Gel Check
Chiba-0825-5.JPG
Sample No 1~3
Sample DNA 2
Loading Dye 2
H2O 8
TOTAL 12
From left;
  1. insert:C0178 -> OK
  2. insert:C0170 -> OK?
  3. vector:J04500 -> OK


--->SAP
Sample No 3
Sample DNA 26
SAP 1
SAP Buffer 3
TOTAL 30μL


--->Zymo Clean
Sample No.3->9μl


--->Gel Check
Chiba-0826.JPG
Sample No.3 1
Loading Dye 1
dH2O 4
TOTAL 6
Sample No.3 -> OK ･･･30ng?


--->Ligation
]

</tr>

Sample No (1)(2)(3)(4)(5)
insert①C0178 1--1-
insert②C0170 -1--1
vector③J04500 --222
ligase 11111
Buffer 22222
dH2O 66544
TOTAL 1010101010


--->Transformation
  1. insert①C0178
  2. insert②C0170
  3. vector③J04500
  4. insert①C0178+vector③J04500
  5. insert②C0170+vector③J04500


27 August, 2008

Digestion Test

Sample DNA : BBa_R0010[39]
Sample No 123
Sample DNA 111
PstⅠ --0.5
EcoRⅠ -0.50.5
Buffer 3 -11
BSA --1
dH2O -7.52
TOTAL 11010


--->Gel Check
Chiba-0827.JPG
Sample No 123
Sample DNA 155
Loading Dye 111
H2O 4--
TOTAL 666
From left;
  1. BBa_R0010
  2. Single Digestion
  3. Double Digestion


(26/8)--->Colony-PCR
Colony PCR of 5 colonies from ligation plates (26/8-(4),(5)) and one from control plate(BBa_F2620[40](2007)).
DNA Template 1
dNTP mix 5
Foward Primer 5
Reverse Primer 5
DNA polymerase 0.5
Thermopol Buffer 5
dH2O 28.5
TOTAL 50μL


95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃



--->Gel Check
Chiba-0827-2.JPG
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)Miniprep


Transformation

competent cells : XL10G
  • BBa_C0062(2007) [41]
  • BBa_C0062(2006) [42] -> no colony on plates
  • BBa_C0179(2007) [43]
  • BBa_C0179(2006) [44]
  • BBa_S03156(2007) [45]
  • BBa_S03156(2006) [46]
  • BBa_S03158(2007) [47]
  • BBa_S03158(2006) [48]
  • BBa_S03160(2007) [49]
  • BBa_S0316(2006) [50]


--->Mini prep


--->(29/8)Gel Check


28 August, 2008

(27/8)--->Mini prep
  1. insert:C0170 + vector:J04500
  2. insert:C0178 + vector:J04500


--->Digestion Test
  1. insert:C0170 + vector:J04500
  2. insert:C0178 + vector:J04500
Sample No 3456
insert:C0170 + vector:J04500 1-1-
insert:C0178 + vector:J04500 -1-1
EcoRⅠ 0.10.10.10.1
SpeⅠ --0.10.1
Buffer 3 1111
BSA --11
dH2O 7.87.86.86.8
TOTAL 10101010


--->(29/8)Gel Check
Chiba-0828.JPG
Sample No 1,23~6
Sample DNA 110
Loading Dye 12
H2O 4-
TOTAL 612
From left;
  1. 　-> OK
  2. 　-> OK
  3. 　-> OK
  4. 　-> OK
  5. 　-> OK
  6. 　-> OK

29 August, 2008

--->Digestion Test
  1. BBa_R0010(2007) [51]
  2. BBa_C0062(2007) [52]
  3. BBa_C0179(2007) [53]
  4. BBa_C0179(2006) [54]
  5. BBa_S03156(2007) [55]
  6. BBa_S03156(2006) [56]
  7. BBa_S03158(2007) [57]
  8. BBa_S03158(2006) [58]
  9. BBa_S03160(2007) [59]
  10. BBa_S03160(2006) [60]


Sample No S1~10W1W2~10
Sample DNA 142
PstⅠ -0.0750.1
EcoRⅠ 0.10.0750.1
Buffer 3 10.751
BSA -0.751
dH2O 7.94.355.8
TOTAL 101010


--->Gel Check
Chiba-0829.JPG
Sample No. 1~10S1~S10,W1~W10
Sample DNA 110
Loading Dye 12
TOTAL 612
From left;
1~5,S1,S5,W1~W5
Chiba-0829-2.JPG
From left;
6~10,S6~S10,W6~W10


(27/8)--->Gel Check
Chiba-0829-3.JPG
Sample DNA 0.5
Loading Dye 1
dH2O 4.5
TOTAL 6μL
R0010 -> OK


Digestion

  1. BBa_I9026(2007) [61]
  2. BBa_I9030(2007) [62]
  3. BBa_S03154(2007) [63]
  4. BBa_R0010(2006) [64]


Sample No 1~34
Sample DNA 100ng/μL×20200ng/μL×30
PstⅠ 22
XbaⅠ 2-
SpeⅠ -2
Buffer 2 -5
Buffer 3 5-
BSA 55
dH2O 166
TOTAL 5050
--->Gel Check
Chiba-0829-4.JPG
Sample No. 1~34
Sample DNA 103
Loading Dye 22
dH2O -7
TOTAL 1212
insert-I9026 : 685bp -> None
Chiba-0829-5.JPG
insert-I9030 : 745bp -> None
Chiba-0829-6.JPG
insert-S03154 : 685bp -> None
Chiba-0829-7.JPG
vector-R0010 : 2079bp -> OK


--->Gel Extract


--->Zymo Clean
->15μL
--->SAP
Sample No 4
Sample DNA 15
SAP 2
SAP Buffer 3
dH2O 10
TOTAL 30μL


--->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

Transformation
competent cells : XL10G
  • BBa_I9026(2007) [65]
  • BBa_I9030(2006) [66]
  • BBa_S03154(2007) [67]
  • BBa_S03156(2007) [68]
  • BBa_S03158(2007) [69]
  • BBa_S03160(2007) [70]
  • BBa_C0062(2007) [71]
  • BBa_C0179(2007) [72]
--->(31/8)Mini prep


(29/8)--->Mini prep
  1. insert:C0170 + vector:J04500
  2. insert:C0178 + vector:J04500


--->Digestion Test
  • insert:C0170 + vector:J04500 -> Sample No.1~4
  • insert:C0178 + vector:J04500 -> Sample No.5~8
  • BBa_T9002(2007)[73] -> 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
Sample No.Single : 10~18Double : 19~27
Sample DNA15
XbaⅠ0.10.1
SpeⅠ-0.1
Buffer 211
BSA11
dH2O6.92.8
TOTAL1010


--->Gel Check
Chiba-0830.JPG
Sample No 1~910~1819~27
Sample DNA 11010
Loading Dye 122
dH2O 4--
TOTAL 61212
From left;
Sanple No.1~13
-> OK
Chiba-0830-2.JPG
From left;
Sample No.14~17,24~16
14 -> OK
15,16 -> Bad
17 -> None
24~16 -> Bad
Chiba-0830-3.JPG
From left;
Sample No.18,27,19~23
18,27 -> Bad
19~22 -> OK
23 -> OK??
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
Retrieved from "http://2008.igem.org/Team:Chiba/jk/%CE%B2/weak_2"