wrong cut size - (Apr/07/2008 )
Sorry, for the stupid question but is the BSA 1x in your final reaction?
I agree with MolBioGirl. ON-incubation might be a bit long. I think you could even try a 2h-incubation. If you would linearize the plasmid in parallel and run both reactions on the gel, you would see whether the small part has been cut out or not.
-zek-
QUOTE (zek @ Apr 7 2008, 03:10 PM)
Sorry, for the stupid question but is the BSA 1x in your final reaction?
I agree with MolBioGirl. ON-incubation might be a bit long. I think you could even try a 2h-incubation. If you would linearize the plasmid in parallel and run both reactions on the gel, you would see whether the small part has been cut out or not.
I agree with MolBioGirl. ON-incubation might be a bit long. I think you could even try a 2h-incubation. If you would linearize the plasmid in parallel and run both reactions on the gel, you would see whether the small part has been cut out or not.
Ooh -- that's a good point, Zek. I didn't notice that the amount of BSA listed was 1µl.
YQLU, check your BSA. It's usually supplied as 100x from NEB. So for your 60µl rxn, the amount added should only be 0.6µl.
-MolBioGirl-