Has anyone done dideoxy sequencing using fluorescent probes?
I am trying to optimize a method to sequence a pure RNA sample. I have been trying to perform the reverse transcription based dideoxy sequencing reaction followed by analyzing on a 12% (19:1) acrylamide in Bio-rad Protean XL setup(20 cm gel).
But the main complication is that we do not have a radiolabelling facility so i am trying to work with a fluorescent labelled probe. We run the gel and view it later as we also do not have a DNA sequence.