Team:HokkaidoU Japan/Notebook/September1
From 2010.igem.org
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 1069 bp plus primer added 300 bp 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: λ/Hind III(2 uL)
- 3: pUC119 (3000 bp): 10 ng/uL
- 4: pSB1C3 (2000 bp): 20 ng/uL
- 5: 1-5A (1000 bp): 80 ng/uL