big dye terminator V3.0
aiyar at ebv.mimnet.northwestern.edu
Thu Aug 23 08:40:20 EST 2001
On Wed, 22 Aug 2001 20:15:11 -0400,
Matt Thomas (mthomas9770 at yahoo.com) wrote:
> I'm also interested in recipes on how to dillute BDT mix (buffer
> composition). I know people dillute it and I hear stories of 1:8 but I can
> never get any reasonable resuslts with less than a 1:2 dillution. So if
> anyone can share I would appreicate it.
How do you dilute it? If you use water, you are diluting out the
buffer and divalent cations required by Taq for DNA polymerisation.
We use a product called HalfBD (from Genpak) and routinely do a
1:8 dilution, with 35 cycles.
I've known others to dilute BD out to greater extents and simply
increase the number of cycles to 50 and beyond.
Ashok Aiyar a-aiyar at northwestern.edu
Department of Microbiology-Immunology office: (312) 503-2524
303 E. Chicago Avenue, WARD 4-123 lab: (312) 503-2542
Northwestern University, Chicago, IL 60611 fax: (312) 503-1339
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