Pick a colony (0.5-1.5 mm of diameter) and transfer to a tube containing 100 µl , vortex. Add the following components to a 0.2 ml tube (on ice): Colony lysate 2.0 µl 10X PCR buffer (Fermentas) 5.0 µl 25 mM 4.0 µl 10 mM 4 dNTPs 1.0 µl Taq Polymerase (5U/µl – Fermentas) 0.5 µl 30 pmol each primer (30 pmol/µl) 1.0 µl (X2) mili-Q 36.5 µl Total volume 50.0 µl Do one cycle of 5 min at 95ºC and proceed with the amplification cycles