'Half BD' big dye diluent

Laura A.Cox lcox at icarus.sfbr.org
Tue Nov 21 09:34:08 EST 2000


On 15 Nov 2000, Kevin Clark wrote:

> Has anyone tried the diluents on the market that you can add to BigDye
> to reduce the amount of sequencing kit you use?
>
> Are they any good?
>
> How little BD kit can you use and does it reduce read lengths?
We are currently runnning 377 machines. We routinely use 2ul big dye in a
10ul reaction. We make our own 2.5x Big Dye buffer (200mM Tris pH 9.0, 5mM
MgCl2). After precipitating the sample, we resuspend in 2ul loading dye and
load 1ul onto the gel for a 96 lane gel. We found no decrease in read
length using this dilution. We have been using this protocol for more than
3 years and consistently get good sequencing results.

Laura Cox

*****************************************************************
Laura A. Cox, Ph.D.
Department of Genetics
Southwest Foundation for Biomedical Research
PO Box 760549
San Antonio, TX 78245-0549
(office) 210-258-9687   (fax) 210-258-9883
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