Team:Chiba/jk/β/weak 2

From 2008.igem.org

(Difference between revisions)
(Week 2)
(Removing all content from page)
 
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>[[Team:Chiba/jk/β|送受信班]]
 
-
==Week 2==
 
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===[[Team:Chiba/jk/β/weak 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>
 
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<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/weak 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]]
 
-
|}
 

Latest revision as of 07:06, 26 September 2008