Can we use less BigDye terminator reagent mix?

Andy Vierstraete andy.vierstraete at rug.ac.be
Mon Jul 5 17:13:16 EST 1999


On 4 Jul 1999 08:37:50 -0700, 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?
>
>- Rocky Ho
>


We use the PCR buffer from Qiagen Taq polymerase to dilute the ready reaction
mix.  You always have buffer left when all of the polymerase is used, so we keep
the rest of the buffer for diluting sequencing reactions.

 mix :
 90 ul 10x buffer
 210 ul water
 100 ul Big dye reaction mix

 We use 4 ul of this mix for sequencing ( 1/2 volume reactions), 0,5 ul primer,
 2,5 ul PCR produkt, 3 ul water.  After precipitation, add 2 ul loading buffer
and load everything.  Works fine for us...

 Greetings

 Andy





More information about the Autoseq mailing list