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Colony PCR

  • Take colony, mix with 500ul of RO water
  • Boiling lysis program
  • Then run PCR using Techne TC-312
  • M13 primers (LacZ region of PUC19) and S16 primers (universal bacteria)
  • COL program. 30 cycles. Annealing is 53C (ideal for S16, 56 would be better for M16). Hold at 4

PCR primer design