ArticleRNA (New York, N.Y.)2014
Simple methods for the 3' biotinylation of RNA.
Article in RNA (New York, N.Y.), 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 5 papers.
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Who cites it
5 citing papers in PubMed, 15 citations in OpenAlex.
- RNA aptamers as tools for the purification and analysis of in vivo assembled ribonucleoproteins.RNA (New York, N.Y.) · 2025Review
- RNA-Ligand Interactions Quantified by Surface Plasmon Resonance with Reference Subtraction.Biochemistry · 2022Article
- Synthesis of modified nucleotide polymers by the poly(U) polymerase Cid1: application to direct RNA sequencing on nanopores.RNA (New York, N.Y.) · 2021Article
- Enzymatic RNA Biotinylation for Affinity Purification and Identification of RNA-Protein Interactions.ACS chemical biology · 2020Article
- Aptahistochemistry in diagnostic pathology: technical scrutiny and feasibility.Histochemistry and cell biology · 2017Review
Corrections and comments
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Authors and funding
2 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Biotinylation of RNA allows its tight coupling to streptavidin and is thus useful for many types of experiments, e.g., pull-downs. Here we describe three simple techniques for biotinylating the 3' ends of RNA molecules generated by chemical or enzymatic synthesis. First, extension with either the Schizosaccharomyces pombe noncanonical poly(A) polymerase Cid1 or Escherichia coli poly(A) polymerase and N6-biotin-ATP is simple, efficient, and generally applicable independently of the 3'-end sequences of the RNA molecule to be labeled. However, depending on the enzyme and the reaction conditions, several or many biotinylated nucleotides are incorporated. Second, conditions are reported under which splint-dependent ligation by T4 DNA ligase can be used to join biotinylated and, presumably, other chemically modified DNA oligonucleotides to RNA 3' ends even if these are heterogeneous as is typical for products of enzymatic synthesis. Third, we describe the use of 29 DNA polymerase for a template-directed fill-in reaction that uses biotin-dUTP and, thanks to the enzyme's proofreading activity, can cope with more extended 3' heterogeneities.
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