'Half BD' big dye diluent

Daniel J.Bolland dbolland at hgmp.mrc.ac.uk
Sat Dec 2 21:11:39 EST 2000



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In article <8uuv9e$prk$1 at mercury.hgmp.mrc.ac.uk>, Kevin Clark
<kevin at hammer.imm.ox.ac.uk> 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?
>
> Thanks
>
> Kevin
>
>
> ---
>
>
Dear Kevin,

We have tried the 'half-term' diluent but found it is far simpler just to
use half the recommended amount of BigDye ready reaction (4ul vs. 8ul) in a
10ul reaction with 1.6 pmol of primer. This does not noticeably reduce read
lengths (I regularly get over 800bp of good quality sequence from a plasmid
or pcr template),

Daniel J. Bolland
Institute of Genetics,
Queen's Medical Centre,
University of Nottingham


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