2 October 2008

Wetlab

• The ligations of various PCR products into AK3 were transformed and single colony PCR was carried out to check for correct ligation. The following PCR products were ligated:
  • EpsE 5' integration site (5'E, ~500bp)
  • EpsE 3' integration site (3'E, ~500bp)
  • Spectinomycin resistance gene (spec,800bp)
• The pSB primers were used on the following cycle:
  • 1 cycle - 95^oC for 30 sec
  • 30 cycle - 95^oC for 30 sec, 60^oC for 45 sec, 72^oC for 1 min

• The following colonies appear to have correct inserts: 5'E1, 3'E2, Spec1