Oligonucleotide staining

Garry P. Larson glarson at ccmail.llu.edu
Wed Sep 27 15:06:40 EST 1995


In article <446bk4$1sm at mserv1.dl.ac.uk>, "Juergen Sasse (Path)"
<js1 at mole.bio.cam.ac.uk> wrote:

> Hello Netters,
> 
> Does anybody have a quick and easy staining method for oligonucleotides 
> (20-50mers) separated by PAGE?
> 
> Thanx a lot for your help,
> Juergen
> 
> email: js1 at mole.bio.cam.ac.uk

First you forgot to say if you wanted to gel puify the oligos or if you
were just checking them for size and/or failure sequences.  If you just
want to check them for size then:
Sure you can buy Stains All but how-bout using methylene blue (100mg/500ml)
in dH20.  Its extremely sensitive (can detect ~0.01 OD in a band).  Stain
gel for 15-30 min in solution, wash extensively with dH20 with multiple
wash changes until the bkgrd. is clear (~2hr) and dry the gel on Whatmann
paper.
If your gel purifying, the UV shadowing technique works well but you need
at least a couple OD's to see the oligo.  Alternatively, you can run
preparative and analytical lanes on the same gel, cut off the analytical
lanes, stain them with methylene blue and lay the gel next to the
preparative lanes and excise the bands at the proper size.

GLarson



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