PCR errors:How many really?

Duncan Clark Duncan at genesys.demon.co.uk
Thu Jun 1 04:00:49 EST 1995

In article: <1995May31.172756.85740 at ucl.ac.uk>  dmcbtch at ucl.ac.uk (Tom Chappell) writes:

> sure they behave identically. (I'm doing some GFP things right now and
> it's fairly easy to tell whether or not the GFP has been messed up in a
> way that is critical to the experiment--it's not green...)

Could PCR and expression of GFP be used as a relative indicator of fidelity? 
There isn't any easily available method to assess fidelity. You can use 
Kunkel's phages.  Very accurate method but it doesn't look the simplest to
set up. There is a French P53/yeast system (published in NAR) that looks
very easy but they refuse to even acknowledge letters and it would be difficult
to reproduce. With GFP it will depend on errors knocking out the activity. If
lack of activity only happens with mutation of a few critical residues then
it will not be good indicator.



My mind's made up. Don't confuse me with the facts!

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