Recently I used dUTP synthesized R6-Tag as R-Primer and a promoter specific primer as F-primer for round 1, and Tag with another promoter specific primer for round 2. UDG 37 degree for 30min. Yet the system hasn't work...Would anyone give me some clues? THX~
Jack M Gallup
UDG degrades all dUTP-containing deoxyribonucleic acids. E.g.: Uracil-DNA Glycosylase (UDG, UNG) catalyzes the hydrolysis of the N-glycosylic bond between uracil and sugar, leaving an apyrimidinic site in uracil-containing single or double-stranded DNA. You've degraded your primers before they have a chance to work.
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