Contribution in an anthology

Appeared in: Small Non-Coding RNAs

Editor list: Rederstorff, M.

Publication year: 2021

Pages: 41-58

ISBN: 978-1-0716-1385-6

eISBN: 978-1-0716-1386-3

DOI Link: https://doi.org/10.1007/978-1-0716-1386-3_5

Title of series: Methods in Molecular Biology

Number in series: 2300

Abstract: 

Successful detection of very small RNAs (tiny RNAs, ~8–15 nt in length) by northern blotting depends on tailored protocols with respect to transfer and immobilization on membranes as well as design of sensitive detection probes. For RNA crosslinking to positively charged membranes, we compared UV light with chemical RNA crosslinking by 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC), using either denaturing or native polyacrylamide gels. We show that northern blot detection of tiny RNAs with 5′-digoxigenin-labeled DNA/LNA mixmer probes is a highly sensitive and specific method and, in our hands, more sensitive than using a corresponding DNA/LNA mixmer probe with a 5′-32P-end-label. Furthermore, we provide a robust protocol for northern blot analysis of noncoding RNAs of intermediate size (~50–400 nt).

Harvard Citation style: Wiegard, J., Schlüter, M., Burenina, O., Kubareva, E., Klug, G., Grünweller, A., et al. (2021) Northern Blot Detection of Tiny RNAs, in Rederstorff, M. (ed.) Small Non-Coding RNAs. New York, NY: Humana, pp. 41-58. https://doi.org/10.1007/978-1-0716-1386-3_5

APA Citation style: Wiegard, J., Schlüter, M., Burenina, O., Kubareva, E., Klug, G., Grünweller, A., & Hartmann, R. (2021). Northern Blot Detection of Tiny RNAs. In Rederstorff, M. (Ed.), Small Non-Coding RNAs (pp. 41-58). Humana. https://doi.org/10.1007/978-1-0716-1386-3_5