A method for validating microRNAs in plants by miR-RACE.
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.
College of Horticulture, Nanjing Agricultural University, Nanjing, Jiangsu Province, 210095, People's Republic of China, fanggg@njau.edu.cn.
Jinggui
J
Fang
Xin
X
Sun
D003001:Cloning, Molecular / Q000379:methods; D015723:Gene Library; D035683:MicroRNAs / Q000235:genetics*; D010944:Plants / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods*; D018749:RNA, Plant / Q000235:genetics; D017423:Sequence Analysis, RNA / Q000379:methods
D035683:MicroRNAs; D018749:RNA, Plant