Re: DNA Seq: PCRs

Glenis Wiebe (uunet!ucalgary.ca!gwiebe@mail.uu.net)
Thu, 17 Jun 1999 15:17:02 -0600

Dear Susan,

In addition to the suggestions already posted, our lab recommends the following:

- alter the amount of template according to the length of the PCR product. We
add 5-10 ng of template for every 100 bp in length.
- gel-purify the primers
- adjust the annealing temperature to 4C below the Tm of the primer, up to a
maximum of 60C

We run 20uL reactions with 4uL BigDye reaction mix and 4uL buffer.

Sincerely,
Glenis

--
Glenis Wiebe, M.Sc.
University Core DNA Services
University of Calgary
Calgary, AB, Canada


phone: (403) 220-4503, fax: (403) 283-4907
web-site: http://www.ucalgary.ca/~dnalab






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