Cloning of the mgtc promoter into the pKSII+ plasmid

Kamil

Tasks:

• Amplification of mgtc and 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).