Chapter title |
A Method for Validating MicroRNAs in Plants by miR-RACE.
|
---|---|
Chapter number | 10 |
Book title |
Plant Gene Silencing
|
Published in |
Methods in molecular biology, February 2015
|
DOI | 10.1007/978-1-4939-2453-0_10 |
Pubmed ID | |
Book ISBNs |
978-1-4939-2452-3, 978-1-4939-2453-0
|
Authors |
Jinggui Fang, Xin Sun, Fang, Jinggui, Sun, Xin |
Editors |
Kirankumar S. Mysore, Muthappa Senthil-Kumar |
Abstract |
miRNA prediction algorithms often fail to predict the accurate location of the mature miRNA in a precursor sequence with nucleotide-level precision. miRNAs-rapid amplification of cDNA ends (miR-RACE) is an efficient method to determine the precise sequences of computationally predicted microRNAs (miRNAs). miR-RACE includes the following steps: miRNA-enriched library preparation, two specific 5'- and 3'-miRNA RACE (miR-RACE) PCR reactions, and sequence-directed cloning. The most challenging step is the two gene-specific primers designed for the two RACE reactions. The miR-RACE protocol is rapid and can be executed and completed in 2-3 days. |
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