Team:Chiba/Calendar-Home/8 September 2008

From 2008.igem.org

(Difference between revisions)
(→10 September, 2008)
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--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
--->'''[[Team:Chiba/protocol/DNA Purification/zymocleam|Zymo Clean]]'''
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===10 September, 2008===
 
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-->(10/9)
 
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:Colony Count
 
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#LuxI
 
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#LuxI with LVA
 
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#Las
 
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#Background(R0010)
 
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#Background(C0261)
 
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Pick colony
 
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'''[[Team:Chiba/protocol/PCR|Colony-PCR]]'''
 
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::Colony PCR of 10 colonies from ligation plate(9/9-(2)) and 3 colonies from ligation plate(9/9-(3)) one from control plate(BBa_F2620[http://partsregistry.org/Part:BBa_F2620](2007)).
 
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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">DNA Template</td>
 
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<td>1</td>
 
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</tr>
 
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<tr>
 
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<td>dNTP mix</td>
 
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<td>5</td>
 
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</tr>
 
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<tr>
 
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<td>Foward Primer</td>
 
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<td>0.3</td>
 
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</tr>
 
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<tr>
 
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<td>Reverse Primer</td>
 
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<td>0.3</td>
 
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</tr>
 
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<tr>
 
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<td>taq DNA polymerase</td>
 
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<td>0.5</td>
 
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</tr>
 
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<tr>
 
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<td>Thermopol Buffer</td>
 
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<td>3</td>
 
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</tr>
 
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<tr>
 
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<td>Nuclease free water</td>
 
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<td>20</td>
 
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</tr>
 
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<tr>
 
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<td>TOTAL</td>
 
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<td>30μL</td>
 
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</tr>
 
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</table>
 
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??
 
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:95℃,5min -> ( 95℃,1min -> 52℃,1min -> 72℃,1min )･･･25cycles -> 72℃,10min -> 6℃
 
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??
 
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-->'''[[Team:Chiba/protocol/gelcheck|Gel Check]]'''
 
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--->(28/8)'''[[Team:Chiba/protocol/DNA Purification/sigma|Miniprep]]'''
 
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elute 50μL
 

Revision as of 07:40, 24 October 2008

>Home | Notebook

7 September 2008 <|> 9 September 2008

Contents

Laboratory work

Team:Input

no work

Team:Communication

-->(7/9)

Gel Extract


--->Zymo Clean
->15μL?

9 September, 2008

-->(6/9)PCR of I9030

--->Gel Check

--->Gel Extract

--->Zymo Clean


Team:Output

SAP

  • [http://partsregistry.org/Part:BBa_R0040 BBa_R0040](ptet)①
  • [http://partsregistry.org/Part:BBa_R0010 BBa_R0010](pLac)⑤
Sample No. ①⑤
DNA(digested sample) 76
SAP/td> 22
SAP Buffer 33
TOTAL 3030
TOTAL 3030

-->37℃ 1hour

-->65℃ 30min

-->DNA purification

-->21μl 6μl

-->Gel check

Sample No. each
DNA(digested sample) 1
Loading Dye 1
dH2O 4
TOTAL 6
  • vector:[http://partsregistry.org/Part:BBa_R0040 BBa_R0040](ptet)① 75ng/μl
  • insert:[http://partsregistry.org/Part:BBa_S03158 BBa_S03158](RBS+RhlR)② 33.6ng/μl
  • insert:[http://partsregistry.org/Part:BBa_S03156 BBa_S03156](RBS+lasR)③ 33.6ng/μl
  • insert:[http://partsregistry.org/Part:BBa_S03160 Ba_S03160](RBS+cinR)④ 38.4ng/μl
  • vector:[http://partsregistry.org/Part:BBa_R0010 BBa_R0010](pLac)⑤ 100ng/μl
  • insert:[http://partsregistry.org/Part:BBa_J52008 BBa_J52008](RBS+rLuc)⑥ 33.6ng/μl
Sample No. ①&②①&③①&④①⑤&⑥⑤
DNA(vector) 333352.1
DNA(insert) 333-3.1-
Ligase 111111
Ligase Buffer(x5) 222222
dH2O 11148.94
TOTAL 101010102010
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