Team:Chiba/Calendar-Home/31 August 2008

From 2008.igem.org

(Difference between revisions)
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'''OUTPUT TEAM'''
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>[[Team:Chiba|Home]] | [[Team:Chiba/Notebook|Notebook]]
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<br>Digestion&Ligation
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<br>R0010(plac)+rluciferase
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[[Team:Chiba/Calendar-Home/30 August 2008|30 August 2008 <]]|[[Team:Chiba/Calendar-Home/1 September 2008|> 1 September 2008]]
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<br>digestion
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<br>DNA 10μl
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==Laboratory work==
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<br>Buffer2(x10)  5μl
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===Team:Input===
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<br>speⅠ2μl
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no work
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<br>PstⅠ2μl
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<br>BSA  1μl
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===Team:Communication===
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<br>dH2O 30μl
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:'''[[Team:Chiba/protocol/transformation|Transformation]]'''
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<br>---------------------
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:Competent Cells : XL10G
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<br>               50μl
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::*[http://partsregistry.org/Part:BBa_K084007 BBa_K084007(Plac+RBS+LasI)]
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<br>->37℃　　3hours
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::*[http://partsregistry.org/Part:BBa_K084008 BBa_K084008(Plac+RBS+RhlI)]
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<br>->SAP    1μl
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 +
::---> We saved these plates with a refrigerator.
 +
 
 +
 
 +
:'''[[Team:Chiba/protocol/digestion|Digestion]]'''
 +
::#[http://partsregistry.org/Part:BBa_I9026 BBa_I9026](2007)
 +
::#[http://partsregistry.org/Part:BBa_I9030 BBa_I9030](2006)
 +
::#[http://partsregistry.org/Part:BBa_S03154 BBa_S03154](2007)
 +
::#[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
 +
 
 +
:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<tr>
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<td width="257">Sample No.</td>
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<td>1~3</td><td>4</td>
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</tr>
 +
<tr>
 +
<td>Sample DNA</td><td>12</td><td>10</td>
 +
</tr>
 +
<tr>
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<td>PstⅠ</td>
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<td>0.2</td><td>0.2</td>
 +
</tr>
 +
<tr>
 +
<td>XbaⅠ</td><td>0.2</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>SpeⅠ</td><td>-</td><td>0.4</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 2</td>
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<td>-</td><td>1.5</td>
 +
</tr>
 +
<tr>
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<td>Buffer 3</td>
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<td>2</td><td>-</td>
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</tr>
 +
<tr>
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<td>BSA</td>
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<td>2</td><td>1.5</td>
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</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O</td>
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<td>3.6</td><td>1.4</td></tr>
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<tr>
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<td>TOTAL</td>
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<td>20</td><td>15</td>
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</tr>
 +
</table>
 +
 
 +
 
 +
:--->(1/9)'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
 +
 
 +
 
 +
:(30/8)--->'''[[Team:Chiba/protocol/DNA Purification/sigma|Mini prep]]'''
 +
 
 +
 
 +
:--->'''[[Team:Chiba/protocol/digestion|Digestion test]]'''
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::*Plac+RBS+RhlI Sample No.2~5
 +
::*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
 +
 
 +
<table width="600" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<tr>
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<td width="257">Sample</td>
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<td>Single Digesiton 2~5</td><td>Double Digestion 2~5</td><td>Single Digestion T9002①</td><td>Single Digesiton T9002②</td><td>Double Digestion T9002①②</td>
 +
</tr>
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<tr>
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<td>Sample DNA</td><td>1</td><td>3</td><td>3</td><td>1</td><td>3</td>
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</tr>
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<tr>
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<td>XbaⅠ</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
 +
</tr>
 +
<tr>
 +
<td>SpeⅠ</td><td>-</td><td>0.1</td><td>-</td><td>-</td><td>0.1</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 2</td>
 +
<td>0.9</td><td>0.8</td><td>0.9</td><td>0.9</td><td>0.8</td>
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</tr>
 +
<tr>
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<td>BSA</td>
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<td>1</td><td>1</td><td>1</td><td>1</td><td>1</td>
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</tr>
 +
<tr>
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<td>dH<sub>2</sub>O</td>
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<td>7</td><td>5</td><td>5</td><td>7</td><td>5</td>
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</tr>
 +
<tr>
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<td>TOTAL</td>
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<td>10</td><td>10</td><td>10</td><td>10</td><td>10</td>
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</tr>
 +
</table>
 +
 
 +
 
 +
:--->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
 +
 
 +
{|align="justify"
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|[[Image:Chiba-0831-p87.JPG]]
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|
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:<table width="315" border="2" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<tr>
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<td width="257">Sample No.</td>
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<td>Plac+RBS+RhlI Sample No.2~5, [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①②
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</td><td>Single & Double Digestion</td>
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</tr>
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<tr>
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<td>Sample DNA</td>
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<td>1</td><td>10</td>
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</tr>
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<tr>
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<td>Loading Dye</td>
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<td>1</td><td>2</td></tr>
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<tr>
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<td>dH<sub>2</sub>O</td>
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<td>4</td><td>-</td>
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</tr>
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<tr>
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<td>TOTAL</td>
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<td>6</td><td>12</td>
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</tr>
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</table>
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:From Left
 +
::*Single Digestion Plac+RBS+RhlI 2~5
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::*Single Digestion [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
 +
::*Double Digestion Plac+RBS+RhlI 2~5
 +
::*Double Digestion [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
 +
::*[http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
 +
|}
 +
 
 +
 
 +
 
 +
 
 +
===Team:Output===
 +
[[Team:Chiba/protocol/digestion|Digestion]]
 +
 
 +
*[http://partsregistry.org/Part:BBa_R0010 BBa_R0010]①
 +
*PCR product([http://partsregistry.org/Part:BBa_J52008 BBa_J52008])②
 +
 
 +
<table width="250" border="4" cellpadding="0" cellspacing="0" bordercolor="#000000">
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<tr>
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<td width="257">Sample No.</td>
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<td>①</td><td>②</td>
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</tr>
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<tr>
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<td>DNA </td>
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<td>10</td><td>10</td>
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</tr>
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<tr>
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<td>SpeⅠ</td>
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<td>1</td><td>0</td>
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</tr>
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<tr>
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<td>PstⅠ</td>
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<td>1</td><td>1</td>
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</tr>
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<tr>
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<td>XbaⅠ</td>
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<td>0</td><td>1</td>
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</tr>
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<tr>
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<td>BSA(x100)</td>
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<td>0.5</td><td>0.5</td>
 +
</tr>
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<tr>
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<td>Buffer2</td>
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<td>5</td><td>0</td>
 +
</tr>
 +
<tr>
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<td>Buffer3</td>
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<td>0</td><td>5</td>
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</tr>
 +
<tr>
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<td>dH<sub>2</sub>O</td>
 +
<td>28.5</td><td>28.5</td>
 +
</tr>
 +
<tr>
 +
<td>TOTAL</td>
 +
<td>50</td><td>50</td>
 +
</tr>
 +
</table>
 +
 
 +
-->37℃ 3hour
 +
 
 +
-->①20μl refine
 +
 
 +
-->②20μl refine

Revision as of 07:25, 24 October 2008

>Home | Notebook

30 August 2008 <|> 1 September 2008

Contents

Laboratory work

Team:Input

no work

Team:Communication

Transformation
Competent Cells : XL10G
  • [http://partsregistry.org/Part:BBa_K084007 BBa_K084007(Plac+RBS+LasI)]
  • [http://partsregistry.org/Part:BBa_K084008 BBa_K084008(Plac+RBS+RhlI)]
---> We saved these plates with a refrigerator.


Digestion
  1. [http://partsregistry.org/Part:BBa_I9026 BBa_I9026](2007)
  2. [http://partsregistry.org/Part:BBa_I9030 BBa_I9030](2006)
  3. [http://partsregistry.org/Part:BBa_S03154 BBa_S03154](2007)
  4. [http://partsregistry.org/Part:BBa_R0010 BBa_R0010](2007)
Sample No. 1~34
Sample DNA1210
PstⅠ 0.20.2
XbaⅠ0.2-
SpeⅠ-0.4
Buffer 2 -1.5
Buffer 3 2-
BSA 21.5
dH2O 3.61.4
TOTAL 2015


--->(1/9)Gel Check


(30/8)--->Mini prep


--->Digestion test
  • Plac+RBS+RhlI Sample No.2~5
  • [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
Sample Single Digesiton 2~5Double Digestion 2~5Single Digestion T9002①Single Digesiton T9002②Double Digestion T9002①②
Sample DNA13313
XbaⅠ0.10.10.10.10.1
SpeⅠ-0.1--0.1
Buffer 2 0.90.80.90.90.8
BSA 11111
dH2O 75575
TOTAL 1010101010


--->Gel Check
Chiba-0831-p87.JPG
Sample No. Plac+RBS+RhlI Sample No.2~5, [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①② Single & Double Digestion
Sample DNA 110
Loading Dye 12
dH2O 4-
TOTAL 612
From Left
  • Single Digestion Plac+RBS+RhlI 2~5
  • Single Digestion [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
  • Double Digestion Plac+RBS+RhlI 2~5
  • Double Digestion [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②
  • [http://partsregistry.org/Part:BBa_T9002 BBa_T9002]①、②



Team:Output

Digestion

  • [http://partsregistry.org/Part:BBa_R0010 BBa_R0010]①
  • PCR product([http://partsregistry.org/Part:BBa_J52008 BBa_J52008])②
Sample No. ①②
DNA 1010
SpeⅠ 10
PstⅠ 11
XbaⅠ 01
BSA(x100) 0.50.5
Buffer2 50
Buffer3 05
dH2O 28.528.5
TOTAL 5050

-->37℃ 3hour

-->①20μl refine

-->②20μl refine

Retrieved from "http://2008.igem.org/Team:Chiba/Calendar-Home/31_August_2008"