lacU169

Karl Fischer tyr-2 at bones.biochem.ualberta.ca
Thu Oct 16 12:43:23 EST 1997


In article <3Q$lVHAWKgR0Ewme at demon.co.uk>, "Dr. Duncan Clark"
<duncan at genesys.demon.co.uk> wrote:

Duncan,

> We use it for fidelity testing polymerases ie 4.5kb pUC containing lacI;
> PCR all the way round using enzyme under test, Dpn I treat (to kill of
> original template), cut and ligate unique RE site in primers and
> electroporate DH5alpha and plate on Xgal, Amp. White colonies are
> wildtype, blues are errors. More blues, the worse the PCR enzyme
> fidelity. Count a few thousand and calculate fidelity.

What strain of E.coli do you use to produce the target template for the
PCR reaction such that it is susceptable to Dpn I digestion (ie. DNA is
dam methylated)?

GM2163?

Cheers

Karl the hepB guy

-- 
Karl Fischer
tyr-2 at bones.biochem.ualberta.ca





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