IUBio

Big Dye Dilution Buffer

Andy Vierstraete andy.vierstraete at rug.ac.be
Wed Apr 21 12:08:30 EST 1999


On 20 Apr 1999 15:48:31 -0700, Mat <mathew at mnightingale.freeserve.co.uk> wrote:

>
>Hi can anyone help me, I am after a supplier of Big Dye Dilution Buffer, or
>alternativley a protocol for making it myself.

We use the PCR buffer from Qiagen Taq polymerase.  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 en
load everything.  Works fine for us...

Greetings

Andy




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