Gradient PCR Pho

Kama

Tasks:

• Amplification of phoP/phoQ

Methods:

• PCR mixture's composition:

  1ul pfu buffer (Fermentas), 1ul MgSO4 (Fermentas), 0,5ul primers, 0,5ul dNTPs (10 mM), 0,25ul pfu turbo polymerase, 0,5ul template DNA from Listeria, optionally: 0,75ul DMSO, solution was topped up with H2O to 10ul.

• PCR programs:

pho

4min 95°C 
 (30s 95°C, 1min 45-55°C, 4min 72°C)x3 
 (30s 95°C, 1min 55-60°C, 4min 72°C)x28 
 10min 72°C 
 ~ 7°C

• Electrophoretic separation on 1% agarose gel

Results: