cloning of PCR products - errors?
Frank O. Fackelmayer
Frank at Fackelmayer.de
Mon Jun 18 08:42:24 EST 2001
"Richard P. Grant" wrote:
> In article <jp9d782uk05.fsf at sirppi.helsinki.fi>,
> Kristo K M Kulju <kulju at sirppi.helsinki.fi> wrote:
> > There is lot of publications about PCR
> > errors but what about cloning procedure? Is is considered to be
> > practically "error free"?
> Yes. That's why it's called 'cloning'.
agreed, it is CONSIDERED to be error free. But of course it is not. Bugs
have numerous ways to ruin your DNA, especially when the DNA contains
repeated elements (leading to recombination) or sequences that encode a
toxic protein. The most common "errors" would be deletions, it´s the
bug´s way of getting rid of disturbing things...
> You could get errors at the ligation sites if you were dead unlucky (but
> it probably wouldn't clone anyway) and any bug that manages to introduce
> errors into a 3 kb plasmid during replication has serious problems . . .
true, based on the error rate of bacterial DNA replication errors are
very unlikely to occur in a plasmid. In single stranded vectors like M13
derivatives, the sequences are much more error prone, however. Again,
deletions are the most likely errors.
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