Category:Protocols/colony-sequence

1 Take 10ul ddH2O into 16 vials seperately. 2 Get colonies from plate with tips,pipet several times in above water and get 3ul into a new PCR tube.

 (16 colonies in total),the 7ul left for culture with LB/antibiotic in 1.5ml tubes.

3 Prepare PCR master mix

 10mM dNTP          0.5ul*17
 T7 Forward primer  0.5ul*17
 DNA Reverse primer 0.5ul*17
 Taq enzyme         1ul*17
 ddH2O              14.5*17
Total               17*17

4 Take 17ul master mix to every PCR tube. 5 PCR 6 Run gel