PCR mgtc, pho

Kama

Tasks:

• Amplification of pho and mgtc

Methods:

• PCR mixture's composition:

  2,5μl pfu buffer (Fermentas) 
  2,5μl MgSO4 (Fermentas) 
  1,5μl primers 
  1μl dNTPs (10 mM) 
  0,5μl pfu turbo polymerase (KNGiE) 
  1μl template (Salmonella)

  The solution was topped up with H2O to 20μl.

• PCR programs:

pho

2min30s 95°C 
 (30s 95°C, 35s 56°C, 3min30s 72°C)x3 
 (30s 58°C, 35s 61°C, 3min30 72°C)x28 
 10min 72°C 
 ~ 7°C

mgtc

1min30s 95°C 
 (30s 95°C, 35s 48°C, 1min 72°C)x3 
 (30s 95°C, 35s 63°C, 1min 72°C)x28 
 10min 72°C 
 ~ 7°C

• Electrophoretic separation on 1% agarose gel

Results:

Notes:

• denaturation temperature value was wrong for pho.