In vitro mutagenesis with rec A
bownja at bham.ac.uk
Tue Jul 11 12:55:33 EST 1995
I am trying to clone a section of a gene (in pAA 121) which carries a single
B.P. substitution into a different vector which carries the same gene of
interest. Restriction analysis suggests that I should be able to perform a
simple double digest, ligate and transform. This routine procedure is used
regularily in our lab, but I am having problems on this occasion e.g.
purifying the similar sized cut vector away from uncut vector.
I am aware of the possibility of cloning in mutations using the characteristic
of rec A to insert ssDNA into homol. regions of dsDNA and I have come across
protocols. I would like to know how widespread the use of this technique is,
whether it is appropiate for my purpose and what pitfalls to watch for.
Thanks in anticipation,
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