A robust and versatile method for production and purification of large-scale RNA samples for structural biology.

A robust and versatile method for production and purification of large-scale RNA samples for structural biology. RNA. 2020 Apr 30;: Authors: Karlsson H, Baronti L, Petzold K Abstract Recent findings in genome-wide transcriptomics revealed that RNAs are involved in almost every biological process, across all domains of life. The characterization of native RNAs, of unknown function and structure, is particularly challenging due to their typical low abundance in the cell and the inherent sensitivity towards ubiquitous RNA degrading enzymes. Therefore, robust in vitro synthesis and extensive work-up methods are often needed to obtain samples amenable for biochemical, biophysical and structural studies. Here, we present a protocol that combines the most recent advances in T7 in vitro transcription methodology with reverse phase ion pairing and ion exchange HPLC purification of RNAs for the production of yield-optimized large-scale samples. The method is easy to follow, robust and suitable for users with little or no experience within the field of biochemistry or chromatography. The complete execution of this method, e.g. for production of isotopically labeled NMR samples, can be performed in less than a week. . PMID: 32354720 [PubMed - as supplied by publisher]
Source: RNA - Category: Genetics & Stem Cells Authors: Tags: RNA Source Type: research