A rapid, quantitative assay for direct detection of microRNAs and other small RNAs using splinted ligation.
The discovery and characterization of microRNAs (miRNAs) and other families of short RNAs has led to a rapid expansion of research directed at elucidating their expression patterns and regulatory functions. Here, we describe a convenient, sensitive, and straightforward method to detect and quantitate specific miRNA levels in unfractionated total RNA samples. The method, based on splinted ligation, does not require specialized equipment or any amplification step, and is significantly faster and more sensitive than Northern blotting. We demonstrate that the method can be used to detect various classes of small regulatory RNAs from different organisms.
Maroney
Patricia A
PA
Center for RNA Molecular Biology and Department of Biochemistry, Case Western Reserve University, School of Medicine, Cleveland, Ohio 44106-4973, USA.
Chamnongpol
Sangpen
S
Souret
Frédéric
F
Nilsen
Timothy W
TW
D001483:Base Sequence; D015152:Blotting, Northern; D006367:HeLa Cells; D006801:Humans; D008024:Ligands; D035683:MicroRNAs / Q000032:analysis* / Q000235:genetics; D015336:Molecular Probe Techniques / Q000706:statistics & numerical data; D009710:Nucleotide Mapping / Q000379:methods* / Q000706:statistics & numerical data; D012680:Sensitivity and Specificity
D008024:Ligands; D035683:MicroRNAs