PCR site-directed mutagenesis

Philipp Wechner philipp.wechner at uibk.ac.at
Fri Jul 19 03:04:28 EST 2002


what do you mean with PCR1 and 2?

amarindc at aol.com schrieb:

> We have been trying to do mutagenesis in certain area of a protein. Three people in my lab are using different set of mutant primers for different aa. mutations. The first PCR goes fine, so apparently does the second. But we are unable to either clone or reamplify the insert using the outside flanking primers (used in PCR 2). We have already tried different conc. of DNA, primers, purification, annealing temp., number of cycles... We have no idea whatsoever on what is going on.
>
> By the way: we have already mutated other aa. in this same region and worked out fine!
>
> Thanx,
>
> Ana
>
> http://biowww.net/mynews/tree.php?group_name=bionet_molbio_methds-reagnts&begin=0
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