PAAG polymerization problem

Hiljar Sibul hsibul at tamm.eenet.ee
Wed Jan 24 10:47:05 EST 1996


Hello,

In the article "Improved separation of PCR amplified VNTR alleles by a
vertical PAAG electrophoresis" (Int J Leg Med 105:355-9) the authors
claim that using pH gradient between the gel and buffer better resolution of
DNA bands is achieved.
They use two types of gels - 6-10% paag as resolving gel and 3%paag as
stacking gel.
In both gels Trizma sulphate is used instead of TBE , pH= 4.5
Also the bisacrylamide content of the gel is lowered ( 1.6% C ).

The authors did not specify the catalysators used for polymerization.
So by default I used 10% APS & TEMED.

While the resolving gel polymerized OK, the stacking one did NOT, even
after de-gasing and adding more catalysators.


Are there any other catalysts that could be used ?

Suggestions welcome.

hsibul at ebc.ee
Hiljar Sibul, Estonian Biocentre



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