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Can we use less BigDye terminator reagent mix?

Paul Shinn pshinn at mail1.sas.upenn.edu
Tue Jul 6 04:37:11 EST 1999


Rocky Ho (RockyHo at cuhk.edu.hk) wrote:
: I am now using 8 µl BigDye terminator reagent mix for 20 µl sequencing
: reaction. Is there any possible suggestion for reducing the use of the
: reagent?

    We routinely do 1/8 BDTerm reactions.  1ul BDT mix + 2ul 5x buffer 
(listed earlier) + DNA + 20pmol primer, and water for a 10ul rxn.  This 
works great for our M13 templates and PCR reactions.  We load 1/2 of this 
on our 96-lane gels and 1/6 to 1/4 of this on our 3700.

---
Paul Shinn    
Sequencing Coordinator                                    ,___o
pshinn at neomorph.bio.upenn.edu                            _-\_<,
Arabidopsis thaliana Genome Center                      (*)/'(*)
http://genome.bio.upenn.edu/ATGCUP.html
(215) 573-7256




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