User:Meng Xiao He/Notebook/fall08/2008/10/24

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PCR flanking regions

Phusion:

  • cbb3III 5
  • cbb3I 3
  • ccoII 5
  • ccoIII 3
  • DuoII 5
  • DuoI 3

35 cycles:

  • 60°C annealing for 5 cycles, 58°C annealing for 30 cycles, 60°C for 5 cycles
  • 1kb extension time


Mix:

  • 2μL 25μM primer pair mix
  • 25μL 2x master mix
  • colony
  • 23μL H2O



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