A single-molecule method for the quantitation of microRNA gene expression Export

@article{citeulike:445690, abstract = {MicroRNAs (miRNA) are short endogenous noncoding RNA molecules that regulate fundamental cellular processes such as cell differentiation, cell proliferation and apoptosis through modulation of gene expression. Critical to understanding the role of miRNAs in this regulation is a method to rapidly and accurately quantitate miRNA gene expression. Existing methods lack sensitivity, specificity and typically require upfront enrichment, ligation and/or amplification steps. The Direct miRNA assay hybridizes two spectrally distinguishable fluorescent locked nucleic acid (LNA)-DNA oligonucleotide probes to the miRNA of interest, and then tagged molecules are directly counted on a single-molecule detection instrument. In this study, we show the assay is sensitive to femtomolar concentrations of miRNA (500 fM), has a three-log linear dynamic range and is capable of distinguishing among miRNA family members. Using this technology, we quantified expression of 45 human miRNAs within 16 different tissues, yielding a quantitative differential expression profile that correlates and expands upon published results.}, author = {Neely, Lori A. and Patel, Sonal and Garver, Joanne and Gallo, Michael and Hackett, Maria and McLaughlin, Stephen and Nadel, Mark and Harris, John and Gullans, Steve and Rooke, Jenny}, citeulike-article-id = {445690}, citeulike-linkout-0 = {http://dx.doi.org/10.1038/nmeth825}, citeulike-linkout-1 = {http://dx.doi.org/10.1038/nmeth825}, day = {20}, doi = {10.1038/nmeth825}, issn = {1548-7091}, journal = {Nature Methods}, keywords = {microarray\_technology, mirna}, month = {December}, number = {1}, pages = {41--46}, posted-at = {2009-11-09 14:49:50}, priority = {1}, publisher = {Nature Publishing Group}, title = {A single-molecule method for the quantitation of microRNA gene expression}, url = {http://dx.doi.org/10.1038/nmeth825}, volume = {3}, year = {2005} }

TY - JOUR ID - citeulike:445690 L3 - citeulike-article-id:445690 N2 - MicroRNAs (miRNA) are short endogenous noncoding RNA molecules that regulate fundamental cellular processes such as cell differentiation, cell proliferation and apoptosis through modulation of gene expression. Critical to understanding the role of miRNAs in this regulation is a method to rapidly and accurately quantitate miRNA gene expression. Existing methods lack sensitivity, specificity and typically require upfront enrichment, ligation and/or amplification steps. The Direct miRNA assay hybridizes two spectrally distinguishable fluorescent locked nucleic acid (LNA)-DNA oligonucleotide probes to the miRNA of interest, and then tagged molecules are directly counted on a single-molecule detection instrument. In this study, we show the assay is sensitive to femtomolar concentrations of miRNA (500 fM), has a three-log linear dynamic range and is capable of distinguishing among miRNA family members. Using this technology, we quantified expression of 45 human miRNAs within 16 different tissues, yielding a quantitative differential expression profile that correlates and expands upon published results. IS - 1 JF - Nature Methods SN - 1548-7091 EP - 46 TI - A single-molecule method for the quantitation of microRNA gene expression PB - Nature Publishing Group VL - 3 SP - 41 KW - microarray_technology KW - mirna AU - Neely, Lori AU - Patel, Sonal AU - Garver, Joanne AU - Gallo, Michael AU - Hackett, Maria AU - McLaughlin, Stephen AU - Nadel, Mark AU - Harris, John AU - Gullans, Steve AU - Rooke, Jenny PY - 2005/12/20/ UR - http://dx.doi.org/10.1038/nmeth825 ER -