hey all
thank you for your help in advance
i want to built Yeast library with mutations only in 2 helixes.
so i have plasmid. in it my gene. now lets say i want to do PCR but enter modifications only in 2 helixes. how would i do it?
thanks
may
use the same way for establishing normal library and it good for you to use cDNA library
There are a variety of methods for doing region specific mutagenesis. To some degree it depends upon the length of the regions you wish to mutagenize. If it is short enough to fit on a single oligo, you could order mixed base oligos to cover the region of interest and then either do PCR or gene synthesis using those olives. Alternately you could use error prone PCR for the regions of interest. If the regions are small enough, just do a series of site specific mutagenesis and change each residue one at a time.
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