do I get right PCR product - (Dec/29/2007 )
QUOTE (phage434 @ Jan 2 2008, 10:55 AM)
That looks a lot better, but the gel lanes are overloaded. Dilute your sample 10x and rerun the gel to get better resolution. You could also run the gel longer to improve resolution where you care. It does look as if you have some primer-dimers being formed at low MW, but these may not matter, depending on what you are doing next.
Incidentally, the black region at the bottom indicates that you are not adding dye to the running buffer, and your EtBr is running toward the negative electrode. You can fix this by adding EtBr to the positive terminal running buffer, or by post-staining.
Incidentally, the black region at the bottom indicates that you are not adding dye to the running buffer, and your EtBr is running toward the negative electrode. You can fix this by adding EtBr to the positive terminal running buffer, or by post-staining.
phage434, the next step I do is cloning my PCR product on a plasmid vector, so I am working on E coli these days, It's also my first time do cloning, do primer-dimers interfere with my cloning?
should I get rid of Taq polymerase and MgCl2 and other PCR reagent in my PCR product?
-Ethan-