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A Novel method to profile miRNA expression in anisms:
Sensitive and specific miRNA qRT-PCR with the NCode™ SYBR® Green miRNA qRT-PCR Kit
Sam An, Mark Landers, Mason Brooks, Pete Joszi, Ranjan J. Perera, and Christopher Adams
Invitrogen Corporation, Carlsbad California
MicroRNAs (miRNAs) are short 22-24 nucleotide RNAs that play a vital roles in regulating gene expression. These single stranded RNA molecules are incorporated into the RISC (RNA-inducing plex) and regulate gene expression through various methods including translational inhibition, transcriptional cleavage and transcriptional gene silencing. Recent interest in these non-coding RNAs has been explosive because of its implication in cellular differentiation and disease genesis. Therefore, there is an urgent need to identify and validate miRNA expression in cell lines, tissues, ans. qRT-PCR has e the standard for microarray data validation, as well as an invaluable tool for quantifying individual or subsets of miRNAs with the greatest sensitivity and accuracy. mercially available miRNA qRT-PCR systems employ proprietary, pre-designed miRNA-specific primers for cDNA synthesis by reverse transcription. Unfortunately, this approach requires that the sequence