Author: Elsie Yekwa; Chutima Aphibanthammakit; Xavier Carnec; Bruno Coutard; Caroline Picard; Bruno Canard; Sylvain Baize; François Ferron
Title: Arenaviridae exoribonuclease presents genomic RNA edition capacity Document date: 2019_2_8
ID: fvp45ho6_4
Snippet: After incubated at 37°C, the reactions were quenched at intervals between 0 and 30 minutes by the addition of an equal volume of loading buffer (formamide containing 10mM EDTA). The products were heated at 70°C for 5 minutes, rapidly cooled on ice for 3 minutes followed by separation in a 20% poly-acrylamide gel containing 8 M urea and buffered with 0,5X Tris-borate-EDTA. Gels were exposed overnight to a phosphor screen and then visualized with.....
Document: After incubated at 37°C, the reactions were quenched at intervals between 0 and 30 minutes by the addition of an equal volume of loading buffer (formamide containing 10mM EDTA). The products were heated at 70°C for 5 minutes, rapidly cooled on ice for 3 minutes followed by separation in a 20% poly-acrylamide gel containing 8 M urea and buffered with 0,5X Tris-borate-EDTA. Gels were exposed overnight to a phosphor screen and then visualized with a phosphoimager FLA-3000 (Fuji). Total RNA degradation products were quantified using Image Guage (Fuji), the speed of cleavage determined and graphs plotted using GraphPad PRISM version 6.0. Experiments were carried out at least in triplicate and only representative gels are shown.
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