Cloning of the mgtc promoter into the pSB1A3 plasmid

Kamil

Tasks:

• Amplification of the mgtc promoter using new primers
• Isolation and digest of the pSB1A3 plasmid

Methods:

• The PCR mix was prepared as follows:

  5μl buffer B (EURx)
  5μl 10mM dNTPs
  5μl forward starter
  5μl reverse starter
  2μl OptiTaq polymerase (EURx)
   2μl Salmonella matrix
   26 μl H2O

• PCR programme:

   4min 95°C 
   (30s 95°C, 35s 56°C, 40s 72°C)x28 
   10min 72°C 
   ~ 4°C
  

• The results were visualised with gel electrophoresis on 1% agarose gel.

Results:

• Nothing

Conclusions:

• The temperature was chosen... poorly.