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Alice Pawlowski 2021. Denaturing RNA electrophoresis in urea polyacrylamide gels (Urea-PAGE) and semi-dry Northern Blot transfer. protocols.io https://dx.doi.org/10.17504/protocols.io.beemjbc6Copy Citation Copied
URL: https://dx.doi.org/DOI:10.17504/protocols.io.beemjbc6
Authors: Alice Pawlowski
Group: Axmann Lab
Summary: RNA is denatured in 8 M Urea and separated in a polyacrylamide gel electrophoresis (PAGE) based on its molecular weight. Using this method, fragments between 2 to 500 nucleotids can be separated. The migration of the RNA depends on the concentration of PAA: the higher the PAA percentage, the better the resolution of low molecular weight fragments and vice versa. We used 10 % PAA to separate fragments between 50 - 200 nucleotides in 10 ml mini-gels (10.5 x 10 cm) using the Hoefer SE260 Mighty Small system. Subsequently, RNA is transfered from the gel to a nylon membrane (Northern blotting) by using aTrans-Blot SD Semi-Dry Electrophoretic Transfer Cell (BioRad).
Affiliations: Institute of Synthetic Microbiology, CEPLAS, Heinrich-Heine Universität, Düsseldorf
Version: 1
Publication Date: 2021
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