beta-agarase

[Paul N Hengen]

Elaine M. Weidenhammer <ew2f+ at andrew.cmu.edu> writes:

>I'm considering purchasing some beta-agarase for purifying DNA from
>agarose gels.  Does anyone have any experience with this product?  I've
>seen the product for sale in both the BMB and US Biochemicals
>catalogues; the latter source appears to be considerably cheaper.  Any
>comments or tips would be appreciated.

I'm also interested in comments about this for recovering DNA from
agarose. Is the beta-agarase the same as "Gelase" produced by
Epicentre Technologies, Madison, Wisconsin?

In this paper,

@article{Serwer1992,
author = "P. Serwer
 and S. J. Hayes
 and E. T. Moreno
 and C. Y. Park",
title = "A Small (58-nm) Attached Sphere Perturbs the Sieving
of 40-80 Kilobase DNA in 0.2-2.5% Agarose Gels: Analysis of
Bacteriophage T7 Capsid-DNA Complexes by Use of Pulsed Field
Electrophoresis",
journal = "Biochemistry",
volume = "31",
pages = "8397-8405",
year = "1992"}

the authors describe the use of "Gelase" with low melt agarose by
first heating the gel slice to 65 C for 10 minutes, lowering the
temperature to 40 C, and adding "Gelase" to digest the agarose.
Does anyone know if this can be done using normal agarose at
1.0 % to 2.0 % at say 37 C without the heat treatment?

Paul N. Hengen
National Cancer Institute
Frederick Cancer Research and Development Center
Frederick, Maryland 21702-1201 USA