Affinity purification of in vitro transcribed RNA is becoming an attractive alternative to purification using standard denaturing gel electrophoresis. Affinity purification is particularly advantageous because it can be performed in a few hours under non-denaturing conditions. However, the performance of affinity purification methods can vary tremendously depending on the RNA immobilization matrix. It was previously shown that RNA immobilization via an optimized λN-GST fusion protein bound to glutathione-Sepharose resin allows affinity purification of RNA with very high purity and yield. This Chapter outlines the experimental procedure employed to prepare the λN-GST fusion protein used for RNA immobilization in successful affinity purifications of RNA. It describes the details of protein expression and purification as well as routine quality control analyses. © 2012 Springer Science+Business Media, LLC.
CITATION STYLE
Di Tomasso, G., Lampron, P., Omichinski, J. G., & Legault, P. (2012). Preparation of λn-GST fusion protein for affinity immobilization of RNA. Methods in Molecular Biology, 941, 123–135. https://doi.org/10.1007/978-1-62703-113-4_10
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