Here's a question.
I'm getting multiple bands from RT-PCR on my gels. I'm using Invitrogen's Superscript one step kit with genomic RNA as my templates. I'm expecting one band in every lane, but I get a lot of minor bands too, some high, some low. For gel 2, I ran a subset where I raised my annealing temps (if I go 2 more degrees higher everything goes away), my extension temps, and my RT temp. Since I'm expecting a 300-400 bp amplicon, I also cut down the time for my DNA extensions to get rid of the big non-specific bands.