Gradient PCR PhoP/PhoQ

Kama

Tasks:

• Amplification of phoP/phoQ

Methods:

• PCR mixture's composition:

  1μl Pfu buffer (EURX) 
  0,25μl primer phoPF
  0,25μl primer phoQR 
  0,5μl dNTPs (10 mM)
  0,25μl Pfu turbo polymerase (EURX)
  0,5μl template DNA from Salmonella (unknown strain)
  0,75μl DMSO 
  The solution was topped up with H2O to 10μl.

• PCR programs:

pho

4min 95°C 
 (30s 95°C, 1min 55-65°C, 5min 10s 72°C)x35
 10min 72°C 
 ~ 7°C

• Electrophoretic separation on 1% agarose gel

Results:

Insertion of the pho gene into the pSB plasmid

Kama

• Isolation of plasmid containing an insert was performed with the A&A plasmid mini kit

Notes