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Team:HokkaidoU Japan/Notebook/aggregation Week 10
From 2012.igem.org
September 3rd
Colony PCR of eCFP-RBS-pSB1A2
| DNA solution | 4 ul |
| Kapa-Taq(Taq polymerase) | 5 ul |
| Forward Primer(ag43-f4 primer) | 0.5 ul |
| Reverse Primer(200bp down primer) | 0.5 ul |
| Total | 10 ul |
| Number | Degree | Second |
| 1 | 95 | 120 |
| 2 | 95 | 30 |
| 3 | 53.0 | 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.
[[image:|thumb|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.
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