question of getting a 75bp PCR product - (Sep/26/2005 )
I want to get a DNA sequece of 75bp through PCR . When using common PCR program, no aimed band showed on 3.0% agrose gel. But it can be dectected out by blotting. How could I alter the PCR program to get it and make it show bright band on agrose gel?
Basically with a product that small you need to load several micrograms on an agarose gel to be able to see it with EtBr staining. Unless you start with PCRs of 100ul and then ethanol precipitate before loading there's nothing you can do with the PCR.
However if you switch to running 20%TBE acrylamide gels (I use the premade NuPage ones from Invitrogen) to make your product visible, you should not have a problem.