A non-radioactive method for small RNA detection by northern blotting
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- 作者：Qi Huang ; Zhinang Mao ; Shaoqing Li ; Jun Hu ; Yingguo Zhu
- 关键词：Small RNA ; Biotin ; labeled probe ; Non ; radioactive ; Northern blot ; Epigenetic
- 刊名：Rice
- 出版年：2014
- 出版时间：December 2014
- 年：2014
- 卷：7
- 期：1
- 全文大小：1,651 KB
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- 出版者：Springer US
- ISSN：1939-8433
文摘
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 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. Conclusions This protocol is an alternative strategy for small RNA detection and represents an efficient means of researching small RNAs.