ArticleRNA (New York, N.Y.)2004
A general method for rapid and nondenaturing purification of RNAs.
Article in RNA (New York, N.Y.), 2004. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 41 papers.
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Who cites it
41 citing papers in PubMed.
- Native RNA Purification Method for Small RNA Molecules Based on Asymmetrical Flow Field-Flow Fractionation.Pharmaceuticals (Basel, Switzerland) · 2022Article
- Sequence-selective purification of biological RNAs using DNA nanoswitches.Cell reports methods · 2021Article
- A non-radioactive, improved PAR-CLIP and small RNA cDNA library preparation protocol.Nucleic acids research · 2021Article
- Single-pass transcription by T7 RNA polymerase.RNA (New York, N.Y.) · 2020Article
- Probing RNA-Protein Interactions and RNA Compaction by Sedimentation Velocity Analytical Ultracentrifugation.Methods in molecular biology (Clifton, N.J.) · 2020Article
- NMR characterization of RNA small molecule interactions.Methods (San Diego, Calif.) · 2019Review
- Coupling Green Fluorescent Protein Expression with Chemical Modification to Probe Functionally Relevant Riboswitch Conformations in Live Bacteria.Biochemistry · 2018Article
- Isotope labeling for studying RNA by solid-state NMR spectroscopy.Journal of biomolecular NMR · 2018Review
- A practical approach to generate suitable de novo synthesis RNA template for a flavivirus RNA-dependent RNA polymerase.Virologica Sinica · 2017Article
- Rapid NMR screening of RNA secondary structure and binding.Journal of biomolecular NMR · 2015Article
- A Platform for Discovery and Quantification of Modified Ribonucleosides in RNA: Application to Stress-Induced Reprogramming of tRNA Modifications.Methods in enzymology · 2015Article
- Native purification and labeling of RNA for single molecule fluorescence studies.Methods in molecular biology (Clifton, N.J.) · 2015Article
- Advances in methods for native expression and purification of RNA for structural studies.Current opinion in structural biology · 2014Review
- RNA synthesis and purification for structural studies.RNA biology · 2014Review
- Strong anion-exchange fast performance liquid chromatography as a versatile tool for preparation and purification of RNA produced by in vitro transcription.RNA (New York, N.Y.) · 2013Article
- Affinity purification of T7 RNA transcripts with homogeneous ends using ARiBo and CRISPR tags.RNA (New York, N.Y.) · 2013Article
- RNA-protein analysis using a conditional CRISPR nuclease.Proceedings of the National Academy of Sciences of the United States of America · 2013Article
- Basis for ligand discrimination between ON and OFF state riboswitch conformations: the case of the SAM-I riboswitch.RNA (New York, N.Y.) · 2012Article
- Ultraviolet shadowing of RNA can cause significant chemical damage in seconds.Scientific reports · 2012Article
- The meaning of a minuscule ribozyme.Philosophical transactions of the Royal Society of London. Series B, Biological sciences · 2011Article
Corrections and comments
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Authors and funding
2 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
A key bottleneck in RNA structural studies is preparing milligram quantities of RNA, and current techniques have changed little in over a decade. To address this, we have developed an affinity tag-based purification method of RNA oligonucleotides. The tag is attached to the 3'-end of almost any desired RNA sequence, allowing for the rapid and specific removal of the RNA of interest directly from in vitro transcription reactions using an affinity column to which a specific RNA-binding protein has been attached. Following a wash, the RNA of interest is eluted by the addition of imidazole to the column, activating a mutant HdeltaV ribozyme incorporated into the tag. The affinity column can then be rapidly regenerated using conditions that release the protein-RNA tag interaction without denaturing the protein. To demonstrate that this method rapidly generates high-quality RNA, we have transcribed, purified, and generated diffraction-quality crystals of a mutant form of the Tetrahymena thermophila P4-P6 domain in a 48-h time period.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.