PCR Using T7 and SP6 Primers

Steven Sullivan sullivan at gwis2.circ.gwu.edu
Mon Aug 4 02:24:37 EST 1997

I use RNA polymerase site primers to do PCR all the time, at anneal
temperatures in the low-mid 50's. Best bet is to have yours custom
synthesized (way cheaper than buying sequencing primers) and making them
longer (I use 23-mers).  Just make sure you check the sequence of the
vector you want to amplify -- Invitrogen's TA vectors, for example, have
different T7 sequence than normal, and many vectors differ in the sequence
downstream of the polymerase site. 

More information about the Methods mailing list