PCR Llo

Kuba

Tasks:

• Amplification of llo(hly) for fusion with the secretion signal peptide

Methods:

• PCR mixture's composition:

  2,5ul pfu buffer (Fermentas), 2,5ul MgSO4 (Fermentas), 1,5ul primers, 1,5ul dNTPs (10 mM), 0,5ul pfu turbo polymerase, 1ul template DNA from Listeria, solution was topped up with H2O to 25ul.

  One of the samples also contained 1 ul of DMSO

• PCR programs:

hly

4min 95°C 
 (30s 95°C, 40s 43°C, 1min30s 72°C)x3 
 (30s 95°C, 40s 45°C, 1min30s 72°C)x28 
 10min 72°C 
 ~ 7°C

random program

• Electrophoretic separation on 1% agarose gel

Results: