Cloning PCR products

Pamela Norton BI599023 at BROWNVM.BROWN.EDU
Wed Jan 29 12:51:55 EST 1992


    While adding extra bases to flank restriction sites in PCR primers is
likely to be a good strategy, the ability of different enzymes to cut near
a free end seems to vary dramatically. New England Biolabs has a table in their
most recent catalogue (p.132 in mine) listing the efficiency of cleavage
near ends for several enzymes. Some cleave very poorly even with four
extra bases; these would probably represent poor choices for primer incorpor-at
ation.

    I hope this helps. I would be interested in hearing about other
compiliations of this type of information which people may have come across.

    Pamela Norton                     bi599023 at brownvm.brown.edu
    Roger Williams Hospital and Brown University
    Providence, RI



More information about the Methods mailing list