Team:HokkaidoU Japan/Notebook/September1
From 2010.igem.org
(Difference between revisions)
(→1-5A(RFP reporter)のPCR) |
(→PCR of 1-5A(RFP reporter)) |
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| 1 uL | | 1 uL | ||
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- | |style="text-align:right;"| Template (1-5A) : | + | |style="text-align:right;"| Template ([[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|1-5A]]) : |
| 1 uL | | 1 uL | ||
|- | |- | ||
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* 3: pUC119 (3000bp): 10 ng/uL | * 3: pUC119 (3000bp): 10 ng/uL | ||
* 4: pSB1C3 (2000bp): 20 ng/uL | * 4: pSB1C3 (2000bp): 20 ng/uL | ||
- | * 5: 1-5A (1000bp): 80 ng/uL | + | * 5: [[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|1-5A]] (1000bp): 80 ng/uL |
Revision as of 07:33, 1 October 2010
PCR of 1-5A(RFP reporter)
Reagent | Amount |
Autoclaved DW : | 33 uL |
10x PCR buffer : | 5 uL |
2 mM dNTPs : | 5 uL |
25 mM MgSO4 : | 3 uL |
100 bp up primer : | 1 uL |
200 bp down primer : | 1 uL |
KOD plus Neo : | 1 uL |
Template (1-5A) : | 1 uL |
Total : | 50 uL |
- Because part lenght was 1069bp plus primer added 300bp to that, the extension time was 90 sec
- PCR product was purified via Microcon YM-10
Electrophoresis
Electrophoresed pUC119,pSB1C3,1-5A to check concentration (1 uL of each)
- 2: Lambda/Hind III(2 uL)
- 3: pUC119 (3000bp): 10 ng/uL
- 4: pSB1C3 (2000bp): 20 ng/uL
- 5: 1-5A (1000bp): 80 ng/uL