zxmkr08 at studserv.zdv.uni-tuebingen.de
Mon Feb 15 13:46:03 EST 1993
In <C2At2B.GAM at ncifcrf.gov> pnh at fcsparc6.ncifcrf.gov (Paul N Hengen) writes:
>In this paper,
>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
>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?
Epicentre says that Gelase can only digest LMP agarose. Maybe they would
be happy if you'd work out conditions for ordinary agarose :-) The heat
treatment is necessary to melt the agarose; Gelase will fail to work
on solid agarose because of sterical reasons (that's again Epicentre).
The high digest temperature leaves the agarose in the molten state (in
newer protocols, the German distributor of Gelase recommends even 45 C
My experiences with Gelase are quite good, BTW.
Hope that helps, Cornelius.
/* Cornelius Krasel, Department of Physiological Chemistry, U Tuebingen */
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