Robust, Linear miRNA Amplification
Most microarray platforms require 1–10 µg of starting total RNA or the equivalent of enriched miRNA. The NCode™ miRNA Amplification System is a highly robust and efficient system for amplifying senseRNA molecules from minute quantities of purified miRNA to generate sufficient amounts of material for downstream research. The system provides consistent and accurate ≥1000-fold amplification of miRNA. The resulting amplified miRNA is in the sense orientation, for direct compatibility with miRNA probe sequences on microarrays. Amplified senseRNA is ideal for expression profiling from very small amounts of starting material because it preserves the relative abundance of the different quantities across experiments. This system is designed for use with up to 30 ng of miRNA (isolated from as little as 50 ng of total RNA) as starting material.
Using the NCode™ miRNA Amplification System, you can expect:
- Increased sensitivity with consistent ≥1,000-fold amplification of miRNA
- Amplified product that faithfully represents the initial miRNA sample (Figure 1)
- Reproducibility with precise amplification from reaction to reaction
Figure 1. Sensitive and accurate miRNA microarray profiling from small samples
Total RNA from large-cell carcinomas and adjacent normal tissue was isolated from multiple patients and enriched for miRNA with the PureLink™ miRNA Purification Kit. Enriched miRNA from 300 ng of total RNA was amplified using the NCode™ miRNA Amplification System and labeled for array analysis with the NCode™ miRNA Labeling System. Additionally, the miRNA from 10 µg of total RNA from the same source was labeled with the NCode™ miRNA Labeling System without amplification and hybridized to NCode™ Multi-Species miRNA Microarrays for analysis. The ratios of miRNA expression in tumor samples vs. adjacent normal tissues were calculated for all human miRNA.