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 in | Biology methods and protocols Vol. 9; no. 1; p. bpae033 |
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| Main Authors | , , |
| Format | Journal Article |
| Language | English |
| Published |
England
Oxford University Press
2024
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| Online Access | Get full text |
| ISSN | 2396-8923 2396-8923 |
| DOI | 10.1093/biomethods/bpae033 |
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| Abstract | 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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| AbstractList | 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.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. 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. 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. 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 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. |
| Author | Li, Zixian Nagar, Bhushan Bilic, Mia |
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| Cites_doi | 10.1126/science.1132505 10.1261/rna.1862210 10.1016/j.bbagrm.2018.09.011 10.1073/pnas.1612444114 10.1371/journal.ppat.1003610 10.1038/nrg3722 10.1016/j.bmc.2012.11.043 10.1016/B978-0-12-420037-1.00017-8 10.1002/2211-5463.12644 10.1261/rna.075697.120 10.1107/S0907444911001314 10.1007/978-3-319-42059-2_1 10.1007/s10858-009-9375-2 10.1038/ni.2048 10.1126/science.1163433 10.1107/S0907444904019158 10.1016/S0092-8674(02)00873-5 10.1093/nar/gng082 10.1038/nrg2172 10.1038/nrg3606 10.1002/chem.201502844 10.1093/nar/gnf055 10.1089/jir.2016.0092 10.1002/wrna.1269 10.1016/j.str.2019.11.017 10.1073/pnas.1412842111 10.1021/bi00411a012 10.1261/rna.056614.116 10.1261/rna.038117.113 10.1016/S0021-9258(19)77302-5 |
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| Keywords | in vitro transcription capping structural biology short RNA chromatography |
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| Title | Isolation of short RNAs with homogeneous 3′-ends using quaternary-amine anion exchange chromatography |
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