Team:HokkaidoU Japan/Notebook/September1
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=1-5A(RFP reporter)のPCR= | =1-5A(RFP reporter)のPCR= | ||
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Revision as of 15:01, 18 September 2010
1-5A(RFP reporter)のPCR
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 |
- パーツは1069bp,primerは300bpのため,余裕を持ってExtensionは90秒で行った
- 得られたPCR solutionをMicrocon YM-10でろ過した
電気泳動
pUC119,pSB1C3,1-5Aの濃度を測定するため,電気泳動した(各1 uL使用)
- 2: λ/Hind III(2 uL)
- 3: pUC119 (3000bp): 10 ng/uL
- 4: pSB1C3 (2000bp): 20 ng/uL
- 5: 1-5A (1000bp): 80 ng/uL