help: primer restriction sites
Mon Sep 4 23:15:51 EST 1995
In article <424pki$rhl at grandcanyon.binc.net>,
movatech at mailbag.com says...
>Frank Chen <yatsen at wam.umd.edu> wrote:
>> I designed a pair of primers which have restriction
>> their 5' ends. I did not add additional bp since I intended
>> that into a PCR vector first. I am wondering if I can
>> them with the enzymes for cloning into an expression vector.
>> appreciate your help if you have done that for these two
>> The enzymes are: Bam HI and Hind III.
>Basically, all enzymes seem to require at least some space
>their site and the end of a PCR product for efficient cutting.
>is one of the best cutters, but it still needs 3-6 base pairs.
>heard that in some cases, three is enough, but I'd recommend
>it 5-6 if you can. I would seriously doubt that you would get
>digestion if the site is right at the end of the product
>JLyon at Novagen.com
There is a table about this problem in the New England Biolabs
Catalog, which lists how much in % of the enzymatic efficiency
you still have with n additional bases...
Children's Hospital, Boston
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