help: primer restriction sites

William Alexander ALEXANDERW at cber.cber.fda.gov
Wed Aug 30 06:08:50 EST 1995


In Article <Pine.ULT.3.91.950830091955.7013A-100000 at rac6.wam.umd.edu>, Frank
Chen <yatsen at wam.umd.edu> wrote:
>Dear Colleagues,
>        I designed a pair of primers which have restriction sites at 
>their 5' ends.  I did not add additional bp since I intended to clone 
>that into a PCR vector first.  I am wondering if I can directly digest 
>them with the enzymes for cloning into an expression vector.  I would 
>appreciate your help if you have done that for these two enzymes.
>        The enzymes are: Bam HI and Hind III.
>
>Frank Chen
>
Check out the tables in the 1995 New England Biolabs catalog: 
p. 208; Cleavage close to the end of DNA fragments (oligonucleotides)
p. 209; Cleavage close to the end of DNA fragments (linerized fragments)
A quick inspection of the second table indicates you may be OK with Bam HI
and you seem to be in trouble with Hind III.  I haven't done the experiment,
since I always make primers with at least 3 extra 5-prime nucleotides. 
These charts may also be available on the WWW on NEB's site also by e-mail: 
info at neb.com
See ya,                                        //\         /\\
Bill Alexander                                || + \ . . / + ||
ALEXANDERW at cber.cber.fda.gov                   \\____\X/____//
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