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

ABSTRACT:

Background

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.

Findings

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 ?g 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.

Conclusions

This protocol is an alternative strategy for small RNA detection and represents an efficient means of researching small RNAs.

SUBMITTER: Huang Q

PROVIDER: S-EPMC4884002 | biostudies-literature | 2014 Dec

REPOSITORIES: biostudies-literature

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

Huang Qi Q Mao Zhinang Z Li Shaoqing S Hu Jun J Zhu Yingguo Y

Rice (New York, N.Y.) 20141001 1

<h4>Background</h4>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.<h4>Findings</h4>In this study, we improved the Northern blot method by using biotin-labeled probes. miRNAs and siRNAs derived from both Arabidopsis thal ...[more]

PMID: 26224555

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