problems with reverse primer

Tiejun Zhao tizhao at UTMB.EDU
Thu Feb 26 10:11:05 EST 1998


Dear Dorian Winter:
        There are two hairpin loop in your reverse primer. Their Tm are
61.1 C and 51.3 C. I think this is the reason of your result.

Tiejun Zhao, Ph.D
Universit of Texas Medical Branch

Mag. Dorian Winter wrote:

> Here´s one for you PCR-gods out there....
>
> I want to amplify a cDNA sequence of about 1000 bp.
> The forward primer is
> 5´-AAGCTTATGGCCATGCTCCTGGGG-3´
> The reverse primer is
> 5´-GCCGACACCAGGTCGACTAAGCTT-3´
>
> I don´t get any product . The forward primer works ok as checked with
> other, internal primers, but I can´t get the reverse primer to work.
> I can´t change the location very much, as it contains a stop codon I
> need.
>
> I would be grateful for any brilliant idea what might be the matter or
> how I could solve the problem.
>
> Dorian Winter






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