URGENT - Primer for sequencing

Klaus Lun zxmdx05
Wed Sep 6 10:09:07 EST 1995


Is there a special programm to desing sequencing-primers ? I am looking for a
point-mutation in a ( already known ) sequence, and I designed the primers
manually ( that means i just copied the sequence for the 20mer oligo ). But
some of them seem not to work; so I wanted to know if there are any special
Parameters that have to be considered especially when designing primers for
sequencing, or if the programms for PCR-Primer design can be used; the fragment
to be sequenced is aprox. 1.2 Bp long, and cloned into the pCR-Vector from
Invitrogens TA-Cloning Kit - Has anybody an idea where I can find the sequence
of this vector ( to check the primers for mispriming in the vector ).
I am using the A.L.F. Sequencer from Pharmacia with unlabeled Primers using the
F*-dATP labelling mix. 
I really would appreciate any help ... :))

Thanks, Klaus




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