sequence is blurry

[Jacqui Whiteway]

gc (genecutl at mendel.berkeley.edu) wrote:
> I've been having trouble reading my sequencing gels because the
> bands tend to get blurry after about 60-100 bases from the bottom
> and I can't tell if I'm looking at single bands or doublets.  I've 
> been running 5% acrylamide gels at 35-40 watts.  The buffer is 
> 1xTBE, except that I add sodium acetate to 1M in the bottom buffer
> after the gel has run about half way.  Any suggestions on sharpening 
> my bands would be grealy appreciated.

> --gc
> TheRoadToNowhere
> http://sp1.berkeley.edu/

We generally use an 8% gel run at 60 Watts and get clear bands.

Jacqui