Simple modifications of the standard DNA sequencing protocol allow for sequencing through siRNA hairpins and other repeats.

摘要: RNAi is a relatively new but powerful technology used for monitoring gene silencing in many species. The RNA corresponding to the of interest can be delivered into cells using various protocols, approach where both sense and antisense strands are located same transcript seems most promising. However, DNA sequencing step challenge due formation strong hairpin structure. In this work we present very simple modification standard protocol that allows through such difficult regions. does not require any additional enzymatic or chemical manipulations. It simply adds 5-min heat-denaturation at 98°C, prior addition Taq mix. inclusion effective different categories templates. Other examples implementation have been described previously more detailed information on plasmid DNAs has presented.