Team:HokkaidoU Japan/Notebook/aggregation Week 10

From 2012.igem.org

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(3rd colony-pcr)
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|5 ul
|5 ul
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|Forward Primer(ag43-f4 primer)
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|Forward Primer(EX-F primer)
|0.5 ul
|0.5 ul
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|Reverse Primer(200bp down primer)
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|Reverse Primer(PS-R down primer)
|0.5 ul
|0.5 ul
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|3
|3
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|68.9
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Desired product is about 776bp.
Desired product is about 776bp.
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[[image:|thumb|Colony PCR result]]
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[[image:HokkaidoU2012 120903 colop eCFP-RBS-pSB1A2 EX-F PS-R (2).jpg|thumb|Colony PCR result]]

Revision as of 22:14, 2 September 2012

September 3rd

Colony PCR of eCFP-RBS-pSB1A2

DNA solution 4 ul
Kapa-Taq(Taq polymerase) 5 ul
Forward Primer(EX-F primer) 0.5 ul
Reverse Primer(PS-R down primer) 0.5 ul
Total 10 ul


Number Degree Second
1 95 120
2 95 30
3 68.9 30
4 72 60
5 72 60
6 4 HOLD

Cycle:2~4 x 35

We used N1 (DW only) and N2 (ptetR-RBS-eCFP-dT-pSB1A2) as controls. Desired product is about 776bp.

Colony PCR result


We confirmed that about 70% of ligated DNA formed our desired construct. We selected No.2 and 5 colony for incubation and store No.7 and 8 colony mixture at 4C. </div></div>


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