PAGE and in-gel ethidium bromide - (Jun/26/2006 )
Hi, I wanted to ask, is there a reason why EtBr can't be added to polyacrylamide gels like in agarose electrophoresis?
I'm running DNA PAGE and I don't have any visible marker to see where my bands are. I've never seen a protocol, using EtBr in polyacrylamide gels, only after-run staining.
Have any of you give it a try?
I'm running DNA PAGE and I don't have any visible marker to see where my bands are. I've never seen a protocol, using EtBr in polyacrylamide gels, only after-run staining.
Have any of you give it a try?
I think the main reason is that it would be a mess: In our lab the PAGE devices are also used for protein gels and so on and these gels should be free of EtBr.
Hobglobin
Agreed. I'd keep the EtBr out of the PAGE apparatus if you can help it. It'd work if you added to the gel (not to the samples, the EtBr will run in the other direction and the stain will be lame) but like hobgoblin suggested it's a mess. Best to keep EtBR contained to dedicated apparatuses--I'd go with the post-run stain, since I suspect your PAGE equipment is used for protein most of the time.
It is used for proteins most of the time. Thanks all.
I'm running DNA PAGE and I don't have any visible marker to see where my bands are. I've never seen a protocol, using EtBr in polyacrylamide gels, only after-run staining.
Have any of you give it a try?
EtdBr inhibits polymerisation of acrylamide and thus cannot be used directly when casting the gel (ref. Sambrook, 2000).