Search found 6 matches

by buthercup
Tue Aug 03, 2010 7:03 pm
Forum: Molecular Biology
Topic: separation problem with GelRed
Replies: 12
Views: 33842

Hi,
I tried also SYBR safe and I'm happier with GelRed...,
any way, I really encourage you to change the concentration fo your running buffer from 1X to 0.5X.
by buthercup
Tue Aug 03, 2010 6:57 pm
Forum: Molecular Biology
Topic: plasmid preparation
Replies: 1
Views: 1219

plasmid preparation

Hi everyone, I'm working with Gram-negative bacteria, trying to charactirize some strains isolated form nature. After succesfull DNA extraction with a comertial kit (GeneElute Bacterial Genomic DNA, Sigma), does anyone know how can I verify if the genomic DNA (total DNA) preparation contains also pl...
by buthercup
Thu Mar 04, 2010 12:55 pm
Forum: Molecular Biology
Topic: separation problem with GelRed
Replies: 12
Views: 33842

Hi! I've been using GelRed for a while, after using EtBR extensivelly, and I didn't notice any change or problem with the separation of the bands... what gel % are you using? Im my lab, we usually use 0.8% agarose, TAE1X to disolve agarose, but 0.5X as running buffer, 100V. Adding the dye to the aga...
by buthercup
Thu Mar 04, 2010 12:42 pm
Forum: Molecular Biology
Topic: SDS-PAGE migration conditions
Replies: 4
Views: 3714

Re: SDS-PAGE migration conditions

But of course the gel % have an influence... It's say the equation for the Joule Heat: P (heat) = V * I = V^2/R = I^2 * R If the % is lower..., then the resistance (R) is lower..., then if I don't want to burn the gel during migration, I have to change (maybe just a little) the migration conditions....
by buthercup
Thu Mar 04, 2010 9:53 am
Forum: Molecular Biology
Topic: SDS-PAGE migration conditions
Replies: 4
Views: 3714

SDS-PAGE migration conditions

Hello every body, Hope someone could help me. I'm trying SDS-PAGE for visualisation of polysaccahrides, and I wanted to try different % of polyacrylamide. Working with protein, I' usually use 12% gels, in a continuous system buffer (Laemmli buffer), and I use 20mA/gel. To separate polysaccharide, th...
by buthercup
Fri Feb 13, 2009 12:28 am
Forum: Molecular Biology
Topic: problems with electrophoresis
Replies: 1
Views: 1973

problems with electrophoresis

Hi every one I hope some people could help me. I'm quite disappointing and perplexed, because after performing a protein extraction from bacteria, with a typical 2D buffer (Tris, Urea, Thiourea, CHAPS), I quatify the extraction and obtain about 4 ug/ul, quite a big concentration. But in the moment t...