PCR mgtc, hly

Kamila

Tasks:

• Amplification of hly

Methods:

• PCR mixture's composition:

  2,5ul pfu buffer (Fermentas), 2,5ul MgSO4 (Fermentas), 1,5ul primers, 1,5ul dNTPs (10 mM), 01,ul pfu turbo polymerase (Fermentas), 1ul template DNA from Listeria, solution was topped up with H2O to 25ul.

  1 repeat of every sample was made (2 different programs).

• PCR programs:

mgtc

90s 95°C 
 (30s 95°C, 35s 48°C, 60s 72°C)x2 
 (30s 95°C, 35s 58°C, 60s 72°C)x28 
 600s 72°C 
 ~ 4°C

hly

90s 95°C 
 (30s 95°C, 35s 42°C, 150s 72°C)x2 
 (30s 95°C, 35s 47°C, 150s 72°C)x28 
 600s 72°C 
 ~ 10°C

• Electrophoretic separation on 1% agarose gel

Results:

Notes:

• Inappropriate template DNA was used for mgtc (ganomic DNA from Yersinia instead of genomic DNA from Listeria).