messy agarose bands

John Elliott 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.
 jte



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