Dumb Gelase question

[Larry Hale]

I just tried Gelase for the first time, and the recovery was pathetic. Just 
for practice, I tried to recover the 2.32 and 2.03 kb HindIII fragments from 
a digest of 3 ug of lambda DNA, from a 1% LMP-agarose (in TBE) slice 
weighing 66 mg. I figure only 10% of the DNA was recovered.

Are there any really critical steps to this procedure that I should be 
attentive to?

When the precipitation protocols call for "one volume" of 5M Ammonium 
acetate to be added to the Gelased slice, do they mean the volume of the 
original slice alone, or the slice + Gelase buffer? (I'm sorry. It's not 
clear to me)

Is the precipitated DNA pellet particularly prone to loosening from the 
microfuge tube? How careful must one be when decanting the ethanol?

Many thanks for your patience.

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*  Larry Hale                                             *
*    Biology Dept., Univ. of Prince Edward Island         *
*    Charlottetown, P.E.I., Canada                        *
*                                                         *
*  The opinions expressed are not those of UPEI, sadly.   *
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