messy agarose bands
johne at gdpiris.chem.sunysb.edu
Tue Mar 19 23:17:56 EST 1996
In article <4in1pe$hdd at dewey.csun.edu>, "J.C." <hbmth001 at huey.csun.edu> wrote:
> Does anyone know how to make bands in agarose look sharper? When I do
> restriction digests of lambda DNA with Hind III, Bam HI, and Eco RI using
> the respective (NE Biolab) buffers, the Eco RI digest always looks the
> sharpest. The bands in the other digests trail blebs and streaks,
> usually at their ends, often in their middles.
Check your enzyme concentrations (NEB catalog appendex) you only need ~1U/ug DNA
for most cases. Also if these are minipreps, use a phenol/chloroform extraction
before you precipitate the DNA. It makes a big difference on picky enzymes.
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