Annealing temperature for oligos
Thomas at hastingslab.harvard.edu
Wed Nov 22 11:44:03 EST 1995
Adela.Torres at uv.es (Adela Torres Calatayud) wrote:
>I'm currently working on DNA non-radioactive cycle sequencing, and I'd like to
>know if there is a reliable formula by which I can choose the annealing
>temperature of the PCR program when I'm sequencing using custom-made oligos.
>Right now the formula I'm using is:
> 2(A+T) + 4(G+C) - 6 = T ann.
>But it didn't give me good results when I tried it. So if there's anyone out
>there that can help me, thanks very much! E-mail or post.
> Adela Torres
> (Adela.Torres at uv.es)
I don't where you got the formula, the one I am using Is
Tm = 4(G+C) + 2(A+T)
Ref.: Thein and Wallace 1986, Human Genetic Deseases: A practical approach, K. E. Davis, ed., IRL Press
Hope that helps
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