Isolation of short RNAs with homogeneous 3′-ends using quaternary-amine anion exchange chromatography

Visualizing RNA–protein interactions through structural approaches requires the use of RNA molecules purified to homogeneity. We describe here a simple and effective method, free of acrylamide contamination and without using UV radiation, to separate in vitro synthesized, heterogeneous RNA transcrip...

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Published inBiology methods and protocols Vol. 9; no. 1; p. bpae033
Main Authors Li, Zixian, Bilic, Mia, Nagar, Bhushan
Format Journal Article
LanguageEnglish
Published England Oxford University Press 2024
Subjects
Innovations
in vitro transcription
capping
structural biology
short RNA
chromatography
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ISSN2396-8923
2396-8923
DOI10.1093/biomethods/bpae033

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Summary:Visualizing RNA–protein interactions through structural approaches requires the use of RNA molecules purified to homogeneity. We describe here a simple and effective method, free of acrylamide contamination and without using UV radiation, to separate in vitro synthesized, heterogeneous RNA transcripts (up to ∼15 nucleotides) at single-nucleotide resolution by quaternary-amine anion exchange chromatography. The quality of short RNAs isolated through this method is validated by gel electrophoresis, mass spectrometry, and crystallization with a protein-binding partner.
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ISSN:2396-8923
2396-8923
DOI:10.1093/biomethods/bpae033