I have a few of questions concerning 2D SDS PAGE gel electrophoresis (using IPG strips):
What would cause a spot to streak vertically from its origin towards the buffer front?
I seem to get blank areas after silver staining my 2D gels - any ideas to a cause? I use a silver diamine stain with 0.83% sodium thiosulphate in the gel matrix.
What could cause a background 'cloud' near the end of a gel? I have tried reducing the IPG buffer down to 0.5% but still get a smear.
I have read quite a few troubleshooting guides and have not been able to solve the above questions. I am contemplating taking a 2D course. However, the only two I have found are in Europe. Does anyone know of a 2D electrophoresis course in North America?
Michael Johns
Lacombe Research Centre
Agriculture and Agri-Food Canada
6000 C & E Trail
Lacombe, Alberta
Canada
T4L 1W1
Phone: 403-782-8100 ext. 268
Fax: 403-782-6120
johnsm@em.agr.ca
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