Melbourne/Lab BL Notebook/20071005PCR2


PCR reaction to attach prefix and suffix to the PsrfA promoter.

PCR mix PCR program
5ul 10x buffer\\

2.5ul 10x Enhancer

0.6ul dNTPs (25mM stock)

2.5ul MgSO4 (Supplied in PCR kit)

1.5ul Primer BL_psrfA_fwd (10uM stock)

1.5ul Primer BL_psrfA_rev (10uM stock)

1ul Template (pJS34)

0.4ul Pfx Platinum (Invitrogen)

32.9ul ddH2O

94°C - 5'

94°C - 30"

50°C - 35"

68°C - 1' (goto step 2 x30)

68°C -10'

4°C forever

50ul Total

=> No product. Need to play around with conditions.