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Contribution to Book
Identification of microRNAs and other tiny noncoding RNAs by cDNA cloning
RNA Interference, Editing, and Modification (2004)
  • Victor R. Ambros
  • Rosalind C. Lee
Abstract
MicroRNAs (miRNAs) and other small RNAs can be identified by cloning and sequencing cDNAs prepared from the ∼22-nt fraction of total RNA. Methods are described for the construction of cDNA libraries from small noncoding RNAs through the use of T4 RNA ligase, reverse transcriptase, and polymerase chain reaction. cDNAs are cloned in λ or plasmid vectors, and the sequences are compared to annotated genomic sequence databases, and analyzed by RNA folding programs to distinguish miRNA sequences from other small RNAs of similar size. Northern blot hybridization is used to confirm the expression of small RNAs in vivo.
Publication Date
2004
Publisher
Humana Press
Series
Methods in Molecular Biology
ISBN
978-1-59259-775-8
Citation Information
Victor R. Ambros and Rosalind C. Lee. "Identification of microRNAs and other tiny noncoding RNAs by cDNA cloning" RNA Interference, Editing, and Modification Vol. 265 (2004)
Available at: http://works.bepress.com/victor_ambros/52/