Team:HokkaidoU Japan/Notebook/plastic Week 7

From 2012.igem.org

(Difference between revisions)
(Electrophoresis)
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<div class="hokkaidou-notebook-daily">
<div class="hokkaidou-notebook-daily">
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==August 16th==
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==August 16th==
<div>
<div>
==PCR==
==PCR==
<p>
<p>
-
A PCR of pGEM to get linear PhaA and PhaB using Laury primer and X-PhaA-up-F, S-PhaA-dw-R, X-PhaB-up-F, S-PhaB-dw-R.  
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PCR of pGEM to get linear PhaA and PhaB using Laury primer and X-PhaA-up-F, S-PhaA-dw-R, X-PhaB-up-F, S-PhaB-dw-R.  
{|class="hokkaidou-table-digestion"
{|class="hokkaidou-table-digestion"
   |-
   |-
-
   |pGEM(pGEM 1ul + DW 9ul)
+
   |pGEM(pGEM 1 ul + DW 9 ul)
   |1 ul
   |1 ul
   |-
   |-
-
   |up primer(10mM)
+
   |up primer(10 mM)
   |1 ul
   |1 ul
   |-
   |-
-
   |down primer(10mM)
+
   |down primer(10 mM)
   |1 ul
   |1 ul
   |-
   |-
-
   |25mM MgSO4
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   |25 mM MgSO4
   |3 ul
   |3 ul
   |-
   |-
-
   |2mM sNTPs
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   |2 mM sNTPs
   |5 ul
   |5 ul
   |-
   |-
   |KOD plus neo
   |KOD plus neo
-
   |1ul
+
   |1 ul
   |-
   |-
   |10xPCR buffer
   |10xPCR buffer
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   |}
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{|class="hokkaidou-table-pcr-time"
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|-
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PCR:60sec, 30 cycle 1h8m
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|Number
 +
|Degree
 +
|Second
 +
|-
 +
|1
 +
|94
 +
|120
 +
|-
 +
|2
 +
|98
 +
|10
 +
|-
 +
|3
 +
|68
 +
|60
 +
|-
 +
|4
 +
|4
 +
|HOLD
 +
|}
 +
Cycle:2~3 x 30
Did not work well?
Did not work well?
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==August 17th==
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==August 17th==
<div>
<div>
-
 
==Electrophoresis==
==Electrophoresis==
<p>
<p>
-
We confirmed success of PCR of yesterday by electrophoresis.
+
We confirmed success of PCR of August 16th by electrophoresis.
</p>
</p>
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</p>
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</div>
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</div><div>
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<div class="hokkaidou-notebook-daily">
 
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==August  18th==
 
-
<div>
 
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==Calculation of concentration for digestion==
 
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-
 
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</div><div>
 
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Revision as of 15:48, 26 September 2012

Contents

August 16th

PCR

PCR of pGEM to get linear PhaA and PhaB using Laury primer and X-PhaA-up-F, S-PhaA-dw-R, X-PhaB-up-F, S-PhaB-dw-R.

pGEM(pGEM 1 ul + DW 9 ul) 1 ul
up primer(10 mM) 1 ul
down primer(10 mM) 1 ul
25 mM MgSO4 3 ul
2 mM sNTPs 5 ul
KOD plus neo 1 ul
10xPCR buffer 5 ul
DW 33 ul
Number Degree Second
1 94 120
2 98 10
3 68 60
4 4 HOLD

Cycle:2~3 x 30

Did not work well?


August 17th

Electrophoresis

We confirmed success of PCR of August 16th by electrophoresis.

Gel extraction

The plasmids done PCR were extracted from TBE gel, and then we got 50 ul DNA solutions.