Team:Chiba/Calendar-Home/31 August 2008

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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 : XL10Gold
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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>
 +
</tr>
 +
<tr>
 +
<td>Sample DNA(μL)</td><td>12</td><td>10</td>
 +
</tr>
 +
<tr>
 +
<td>PstⅠ(μL)</td>
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<td>0.2</td><td>0.2</td>
 +
</tr>
 +
<tr>
 +
<td>XbaⅠ(μL)</td><td>0.2</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>SpeⅠ(μL)</td><td>-</td><td>0.4</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 2(μL)</td>
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<td>-</td><td>1.5</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 3(μL)</td>
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<td>2</td><td>-</td>
 +
</tr>
 +
<tr>
 +
<td>BSA(μL)</td>
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<td>2</td><td>1.5</td>
 +
</tr>
 +
<tr>
 +
<td>dH<sub>2</sub>O(μL)</td>
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<td>3.6</td><td>1.4</td></tr>
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<tr>
 +
<td>TOTAL(μL)</td>
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<td>20</td><td>15</td>
 +
</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]]'''
 +
::*[http://partsregistry.org/Part:BBa_K084008 BBa_K084008] 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">
 +
<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 T9002(1)</td><td>Single Digesiton T9002(2)</td><td>Double Digestion T9002(1)(2)</td>
 +
</tr>
 +
<tr>
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<td>Sample DNA(μL)</td><td>1</td><td>3</td><td>3</td><td>1</td><td>3</td>
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</tr>
 +
<tr>
 +
<td>XbaI(μL)</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td><td>0.1</td>
 +
</tr>
 +
<tr>
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<td>SpeI(μL)</td><td>-</td><td>0.1</td><td>-</td><td>-</td><td>0.1</td>
 +
</tr>
 +
<tr>
 +
<td>Buffer 2(μL)</td>
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<td>0.9</td><td>0.8</td><td>0.9</td><td>0.9</td><td>0.8</td>
 +
</tr>
 +
<tr>
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<td>BSA(μL)</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(μL)</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(μL)</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>[http://partsregistry.org/Part:BBa_K084008 BBa_K084008] Sample No.2~5, [http://partsregistry.org/Part:BBa_T9002 BBa_T9002](1)(2)
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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(μL)</td>
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<td>1</td><td>10</td>
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</tr>
 +
<tr>
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<td>Loading Dye(μL)</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(μL)</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(μL)</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 [http://partsregistry.org/Part:BBa_K084008 BBa_K084008] 2~5
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::*Single Digestion [http://partsregistry.org/Part:BBa_T9002 BBa_T9002](1),(2)
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::*Double Digestion [http://partsregistry.org/Part:BBa_K084008 BBa_K084008] 2~5
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::*Double Digestion [http://partsregistry.org/Part:BBa_T9002 BBa_T9002](1),(2)
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::*[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)
 +
*[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>
 +
<tr>
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<td>XbaI</td>
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<td>0</td><td>1</td>
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</tr>
 +
<tr>
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<td>BSA(x100)</td>
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<td>0.5</td><td>0.5</td>
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</tr>
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<tr>
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<td>Buffer2</td>
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<td>5</td><td>0</td>
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</tr>
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<tr>
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<td>Buffer3</td>
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<td>0</td><td>5</td>
 +
</tr>
 +
<tr>
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<td>dH<sub>2</sub>O</td>
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<td>28.5</td><td>28.5</td>
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</tr>
 +
<tr>
 +
<td>TOTAL</td>
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<td>50μl</td><td>50μl</td>
 +
</tr>
 +
</table>
 +
 
 +
-->left for 3 hours at 37 °C.
 +
 
 +
-->(1)20μl refine
 +
 
 +
-->(2)20μl refine

Latest revision as of 20:41, 29 October 2008

>Home | Notebook

30 August 2008 <|> 1 September 2008

Contents

Laboratory work

Team:Input

no work

Team:Communication

Transformation
Competent Cells : XL10Gold
  • [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 DNA(μL)1210
PstⅠ(μL) 0.20.2
XbaⅠ(μL)0.2-
SpeⅠ(μL)-0.4
Buffer 2(μL) -1.5
Buffer 3(μL) 2-
BSA(μL) 21.5
dH2O(μL) 3.61.4
TOTAL(μL) 2015


--->(1/9)Gel Check


(30/8)--->Mini prep


--->Digestion test
  • [http://partsregistry.org/Part:BBa_K084008 BBa_K084008] Sample No.2~5
  • [http://partsregistry.org/Part:BBa_T9002 BBa_T9002](1),(2)
Sample Single Digesiton 2~5Double Digestion 2~5Single Digestion T9002(1)Single Digesiton T9002(2)Double Digestion T9002(1)(2)
Sample DNA(μL)13313
XbaI(μL)0.10.10.10.10.1
SpeI(μL)-0.1--0.1
Buffer 2(μL) 0.90.80.90.90.8
BSA(μL) 11111
dH2O(μL) 75575
TOTAL(μL) 1010101010


--->Gel Check
Chiba-0831-p87.JPG
Sample No. [http://partsregistry.org/Part:BBa_K084008 BBa_K084008] Sample No.2~5, [http://partsregistry.org/Part:BBa_T9002 BBa_T9002](1)(2) Single & Double Digestion
Sample DNA(μL) 110
Loading Dye(μL) 12
dH2O(μL) 4-
TOTAL(μL) 612
From Left
  • Single Digestion [http://partsregistry.org/Part:BBa_K084008 BBa_K084008] 2~5
  • Single Digestion [http://partsregistry.org/Part:BBa_T9002 BBa_T9002](1),(2)
  • Double Digestion [http://partsregistry.org/Part:BBa_K084008 BBa_K084008] 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)
  • [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 50μl50μl

-->left for 3 hours at 37 °C.

-->(1)20μl refine

-->(2)20μl refine

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