So, I’ve been doing DNA extractions of planktonic microbial samples, and biofilms…running PCR and then a gel as you do. I’ve seen amazing bands from my reference ladders, but absolutely nothing from my samples unfortunately. Now the reason why has somehow eluded me until now. Ok, Candida is different to bacterial samples, and is a bit more difficult to actually get at the DNA, but surely I should have been getting something to amplify!?
Well, I’ve been changing the variables bit by bit over the past few months, MgCL2 concentration, new and old primers, primer concentrations etc…but yesterday I changed the polymerase. I’ve been using the new G2 Taq polymerase from Promega, but for some reason haven’t been getting any results..however, I switched back to the original Taq and boom, clear bands!!
On the other hand, good progress has been made with the clinical samples – we’re basically ready to go (when I get this DNA extraction sorted at least!!). Exciting times ahead in the very near future 😀