big dye terminator V3.0

Ashok Aiyar 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
-- 
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




More information about the Methods mailing list