Section 58
Chapter 57,079

A non-radioactive method for small RNA detection by northern blotting

Huang, Q.; Mao, Z.; Li, S.; Hu, J.; Zhu, Y.

Rice 7(1): 26


ISSN/ISBN: 1939-8425
PMID: 26224555
DOI: 10.1186/s12284-014-0026-1
Accession: 057078096

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Small non-coding RNAs are essential regulators of gene expression at the transcriptional and posttranscriptional levels. High-throughput sequencing has revealed thousands of predicted small RNAs; however, only a few of these have been well characterized. Northern blotting is the most convincing method for small RNA validation. In this study, we improved the Northern blot method by using biotin-labeled probes. miRNAs and siRNAs derived from both Arabidopsis thaliana and Oryza sativa were investigated. The results suggest that this improved method is sensitive and efficient, with approximately 5 μg of total RNA being sufficient for detection. Furthermore, long-term storage of probes labeled in this manner is more convenient, less contaminative and degradative compared with traditional probes. This protocol is an alternative strategy for small RNA detection and represents an efficient means of researching small RNAs.

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