5' overhang on PCR primers

Dave Knorr dak at apldbio.com
Thu Jan 30 15:08:21 EST 1997


In article <5clo9i$v2a at ds2.acs.ucalgary.ca> Jillian Johnston,
johnstoj at acs2.acs.ucalgary.ca writes:
>Thomas F. Vogt (tvogt at WATSON.PRINCETON.EDU) wrote:
>: I want to do PCR with a primer that has a 5'overhang of 
>: 50bp of unpaired nucleotides for the creation of an 
>: epitope tag. CAN this be done?

I once stuck a T7 promoter plus a restriction site and a few extra bases
to ensure cutting at the 5' end of a cDNA clone for doing in vitro
transcription work.  I used a 70-mer PCR primer and it worked great!

Dave Knorr



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