A simple method for profiling miRNA expression

Jia-Wang Wang; Jin Q Cheng

doi:10.1007/978-1-59745-339-4_14

A simple method for profiling miRNA expression

Methods Mol Biol. 2008:414:183-90. doi: 10.1007/978-1-59745-339-4_14.

Authors

Jia-Wang Wang¹, Jin Q Cheng

Affiliation

¹ H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.

PMID: 18175820
DOI: 10.1007/978-1-59745-339-4_14

Abstract

Here we describe a simple protocol that uses positively charged nylon membrane dot blot to profile miRNA expression. A library of 515 antisense oligodeoxynucleotides of human and mouse mature miRNAs was synthesized and spotted on GeneScreen Plus membrane using a dot-blot equipment. Total RNA or enriched small molecular weight RNAs (smwRNAs) were enzymatically radiolabeled by poly (A) polymerase and then hybridized to the nylon membrane oligo arrays. The spot signal intensity on the membrane was analyzed using phosphorimaging. This method offers a convenient and economic way to simultaneously detect the expression of hundreds of miRNAs.

Publication types

Review

MeSH terms

Animals
Blotting, Southern / methods
Cell Culture Techniques
Gene Expression Profiling / methods*
Humans
MicroRNAs / genetics*
MicroRNAs / isolation & purification
Nucleic Acid Hybridization
Nylons / chemistry
Oligonucleotide Array Sequence Analysis / methods
Staining and Labeling
Tissue and Organ Harvesting

Substances

MicroRNAs
Nylons