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Sequencing without PCR purification

Joakim Westberg jockew at biochem.kth.se
Sat Mar 20 20:16:16 EST 1999

Dear Joao!

If you use less than 10 pmole of each primer in your PCR, you can take 2 ul
of non-purified PCR-product to your sequencing reaction. We do that
routinely in our lab and it work fine. This is with BigDye-terminators. I
don't know how it works with BigDye-primers.

Best Regards,
Joakim Westberg
Dept. of Biotechnology
Kungl. Tekniska H=F6gskolan
Stockholm, Sweden

>I would like to know if someone has obtained good results with ABI
>BigDye sequencing without purifying PCR products (from excess
>primers and dNTPs)?
>Thank you, Joao
>Joao Alexandrino
>Centro Estudos Ciencia Animal-ICETA
>Unidade de Genetica Animal e Conservacao
>Campus Agrario de Vairao
>R. Monte - Crasto - Vairao
>4480 Vila do Conde, Portugal
>tel. 351 52 662399  fax. 351 52 661780
>email: jmalexan at mail.icav.up.pt
>                 ---
>Joao Alexandrino
>Dept. Zoologia e Antropologia
>Faculdade de Ciencias
>Universidade do Porto
>email: jmalexan at fc.up.pt

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