A simple method for profiling miRNA expression.
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.
Wang
Jia-Wang
JW
H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA.
Cheng
Jin Q
JQ
D000818:Animals; D015139:Blotting, Southern / Q000379:methods; D018929:Cell Culture Techniques; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics* / Q000302:isolation & purification; D009693:Nucleic Acid Hybridization; D009757:Nylons / Q000737:chemistry; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D013194:Staining and Labeling; D020858:Tissue and Organ Harvesting
D035683:MicroRNAs; D009757:Nylons