Stratagene ExSite Mutagenesis Kit

J O Mason jom at holyrood.ed.ac.uk
Fri May 5 06:41:13 EST 2000


I'm having a problem with Stratagene's Exsite Site directd mutagenesis
kit. I'm trying to delete about 500bp from a plasmid. The deletion works
OK, but in all clones, I get a single extra nucleotide inserted between
the two oligo sequences (i.e. in place of the delted sequence).
Stratagene suggest that the Pfu polisinh step may have failed, bit I
have tried several Pfus with the same result. It also happens with 3
different primer pairs. Has anyone seen this before, and how dd you get
round it?




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John Mason 

www.ed.ac.uk/~jom/home_page.html

jom at holyrood.ed.ac.uk
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