Team:HokkaidoU Japan/Notebook/September1

From 2010.igem.org

Revision as of 07:33, 1 October 2010 by Sprkata (Talk | contribs)

Notebook

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

Electrophoresis of purified PCR product

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
Retrieved from "http://2010.igem.org/Team:HokkaidoU_Japan/Notebook/September1"