Cloning Micrornas from Mammalian Tissues.
From: Department of Gastroenterology and Hepatology, Flinders Medical Center, Flinders University School of Medicine, Bedford Park, Australia.
Methods in molecular biology (Clifton, N.J.)
- Publish Date: 2006
- ISSN: 1064-3745
- Volume: 342
- Issue:
- Pages: 189-207
- Medium: Print
- Language: English
- Citation (JAMA): Michael Michael Z, et al. Cloning Micrornas from Mammalian Tissues.. Methods Mol. Biol. 2006;342:189-207
Abstract
MicroRNAs (miRNAs) are ubiquitous regulators of gene expression in plants and animals. Their distinctive structure, as very short RNAs with a 5’-phosphate and 3’-hydroxyl group, has enabled the development of protocols to clone miRNAs. After enrichment of these small molecules by size, serial ligation of adapter oligonucleotides to each terminus allows amplification using reverse transcription (RT)-polymerase chain reaction (PCR). Plasmid cloning of multiple miRNA sequences and subsequent DNA sequence analysis enable both bioinformatic characterization of the various miRNAs and experimental validation of their accumulation in cells.
Mesh Headings (Keywords): Animals, Blotting, Northern, Cloning, Molecular, Computational Biology, DNA, Complementary, Gene Expression Regulation, Humans, MicroRNAs, Plasmids, Reproducibility of Results, Reverse Transcriptase Polymerase Chain Reaction, Sequence Analysis, DNA
Check for Full Text / PubMed Unique Identifier (PMID): 16957376
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