ArticleRNA (New York, N.Y.)2013
Strong anion-exchange fast performance liquid chromatography as a versatile tool for preparation and purification of RNA produced by in vitro transcription.
Article in RNA (New York, N.Y.), 2013. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed, 41 citations in OpenAlex.
- Digital Twin Fundamentals of mRNAACS omega · 2024Article
- HPLC for at-line reaction monitoring and purification improves yield and purity of tRNA.Frontiers in molecular biosciences · 2024Article
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- Isolation of short RNAs with homogeneous 3'-ends using quaternary-amine anion exchange chromatography.Biology methods & protocols · 2024Article
- Anion exchange HPLC monitoring of mRNAFrontiers in molecular biosciences · 2024Article
- Strategies for controlling the innate immune activity of conventional and self-amplifying mRNA therapeutics: Getting the message across.Advanced drug delivery reviews · 2021Review
- A robust and versatile method for production and purification of large-scale RNA samples for structural biology.RNA (New York, N.Y.) · 2020Article
- A Modified In Vitro Transcription Approach to Improve RNA Synthesis and Ribozyme Cleavage Efficiency.Molecular biotechnology · 2019Article
- 6S RNA Mimics B-Form DNA to Regulate Escherichia coli RNA Polymerase.Molecular cell · 2017Article
- Article
- The conserved 3'X terminal domain of hepatitis C virus genomic RNA forms a two-stem structure that promotes viral RNA dimerization.Nucleic acids research · 2015Article
- Nonorthogonal tRNA(cys)(Amber) for protein and nascent chain labeling.RNA (New York, N.Y.) · 2015Article
- A Platform for Discovery and Quantification of Modified Ribonucleosides in RNA: Application to Stress-Induced Reprogramming of tRNA Modifications.Methods in enzymology · 2015Article
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Authors and funding
5 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Here we demonstrate the use of strong anion-exchange fast performance liquid chromatography (FPLC) as a simple, fast, and robust method for RNA production by in vitro transcription. With this technique, we have purified different transcription templates from unreacted reagents in large quantities. The same buffer system could be used to readily remove nuclease contamination from the overexpressed pyrophosphatase, the important reagent for in vitro transcription. In addition, the method can be used to monitor in vitro transcription reactions to enable facile optimization of reaction conditions, and we have compared the separation performance between strong and weak anion-exchange FPLC for various transcribed RNAs, including the Diels-Alder ribozyme, the hammerhead ribozyme tRNA, and 4.5S RNA. The functionality of the purified tRNA(Cys) has been confirmed by the aminoacylation assay. Only the purification by strong anion-exchange FPLC has led to the enrichment of the functional tRNA from run-off transcripts as revealed by both enzymatic and electrophoretic analysis.
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