Measuring Poly-Adenosine Tail Length of RNAs by High-Resolution Northern Blotting Coupled with RNase H Cleavage.
Northern blot
Poly(A)
Poly-adenosine tail
RNase H
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2024
2024
Historique:
medline:
2
11
2023
pubmed:
12
10
2023
entrez:
12
10
2023
Statut:
ppublish
Résumé
The poly-adenosine, or poly(A) tail, plays key roles in controlling the stability and translation of messenger RNAs in all eukaryotes, and, as such, facile assays that can measure poly(A) length are needed. This chapter describes an approach that couples RNase H-mediated cleavage of an RNA of interest with high-resolution denaturing gel electrophoresis and northern blot-based detection. Major advantages of this method include the ability to directly measure the abundance of any RNA and the length of its poly(A) tail without amplification steps. The assay provides high specificity, sensitivity, and reproducibility for accurate quantitation using standard molecular biology equipment and reagents. Overall, the high-resolution northern blotting approach offers a cost-effective means of poly(A) RNA analysis that is especially useful for small numbers of transcripts and comparisons between experimental conditions or time points.
Identifiants
pubmed: 37824066
doi: 10.1007/978-1-0716-3481-3_6
doi:
Substances chimiques
Ribonuclease H
EC 3.1.26.4
RNA
63231-63-0
RNA, Messenger
0
Poly A
24937-83-5
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
93-111Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM105707
Pays : United States
Informations de copyright
© 2024. This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply.
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