Duke University/22 June 2009

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June 22, 2009

LB Agar Plates with Chlorophenicol (CAM), anhydrotetracycline, Nile Red (NR)
stock plate
CAM 50 mg/ul 20 ug/ml
anhyd 2 mg/ml 150 ng/ml
NR 0.25 mg/ml 0.5 ug/ml
  • E: PCR with fragments 1 and 2 and different conditions (decreased annealing and extension times and number of cycles), Gel to check PCR
2x Phusion mix 12.5 ul 
pASKphaA 4.76 ul
phaB 0.60 ul
H2O 7.14 ul 
25 ul

98°C 30s 1x
98°C 10s
70-55°C 10s 
72°C 1m 10s 20x
72°C 5m
4°C infinity 1x
  • R: No band/smear
  • C: Change conditions