Simplified miRNA Fluorescent Labeling and Sensitive Detection
The NCode™ Rapid miRNA Labeling System is a fast and reproducible system to directly label endogenous miRNAs with fluorescent tags. Using this system, miRNAs from total RNA samples are labeled with fluorescent Alexa Fluor® dyes and hybridized to microarrays printed with species-specific antisense miRNA probes. This system has been optimized to ensure sensitive and accurate profiling of miRNA expression patterns from minimal RNA input.
Starting with a 250 ng to 5 µg total RNA sample, miRNAs are tagged directly in a quick and easy-to-use protocol with the NCode™ Rapid miRNA Labeling System (Figure 1). The protocol takes approximately one hour to complete and consists of just two steps prior to hybridization: polyA tailing; and ligation of a fluorescent dendrimer. After 8-16 hour hybridization, the microarrays are ready to scan and analyze. The dendrimer is a branched structure of single and double stranded DNA conjugated with Alexa Fluor® dyes, and delivers approximately 15 fluors to the sample.
The high sensitivity is due the dendrimer signal amplification technology, ensuring maximum signal-to-background ratios, and strong signal correlation for increased sensitivity.
The Ncode™ Rapid miRNA Labeling System is available in both single color (Alexa Fluor® 3) and dual color (Alexa Fluor® 3 and Alexa Fluor® 5) labeling formats.
Using the NCode™ Rapid miRNA Labeling System, you can expect:
- Less time at the bench – a single step hybridization and no miRNA enrichment required
- Increased sensitivity - reliably detects attamolar levels of miRNA (approximately 2-10 copies per cell)
- Experiments completed in a single day using the 8 hour hybridization protocol with dynamic hybridization.
Catalog and Ordering
- NCode™ miRNA Rapid Labeling System (dual color)
- NCode™ Alexa Fluor® 3 miRNA Rapid Labeling System (single color)
Want to learn more about labeling and the miRNA profiling workflow? Visit the Epigenetics Learning Center.
Figure 1. NCode™ Rapid miRNA Labeling System Overview.
*Capture reagents contain 3DNA™ reagent manufactured under license from Genisphere, Inc.