PCR mgtc, hly

Kamila

Tasks:

• Amplification of hly

Methods:

• PCR mixture's composition:

  2ul buffer, 1ul MgCl2, 0,5ul primers, 1,5ul dNTPs (10 mM), 01,ul polymerase, solution was topped up with H2O to 20ul.

  2 repeats of every sample were made.

• 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).