RNase R
Effectively enrich circular RNA (circRNA) for NGS or qPCR. RNase R is a 3′ to 5′ exoribonuclease that selectively digests linear RNA, leaving the more stable circRNA intact and unaffected. Treatment with RNase prior to NGS or qPCR increases sensitivity and improves quantification of circRNA for profiling.
Overview
- 3′ to 5′ exoribonuclease that selectively degrades linear RNA
- Enrichment of circRNA improves NGS or PCR analysis
- Removal of linear RNA from circRNA from synthesis samples
RNase R treatment improves quantification and detection of circRNAs for NGS analysis
| Number of BSJs detected at 4.0 x 107 reads | Number of BSJs detected at 1.6 x 108 reads | |||
| – RNase R | + RNase R | – RNase R | + RNase R | |
| HIPK3 | 15 | 131 | 45 | 480 |
| LINC00632 | 248 | 679 | 965 | 2,695 |
| MTO1 | 0 | 10 | 0 | 48 |
| PLOD2 | 0 | 28 | 19 | 124 |
| SMARCA5 | 34 | 100 | 125 | 407 |
| XPO1 | 24 | 63 | 105 | 294 |
RNase R treatment improved quantification and detection of circRNAs for NGS analysis. Panel A. Circular RNA in eukaryotes is formed by backsplicing, which occurs when a downstream splice donor site is joined to an upstream splice acceptor site, looping RNA into a circle. The unique sequence site, where the two exons are joined in reverse order, is referred to as the back-splicing junction (BSJ), and is unique to circRNAs. Panel B. 100 µg of human genomic RNA was divided into two samples. One sample was treated with RNase R (40 U, 37 °C for 30 min) and one sample was untreated. Following treatment, RNA was purified, and 1 ng of purified RNA was used for RNA-seq analysis. The number of reads containing BSJs from six genes reported to express circRNAs was compared between samples with and without RNase R treatment. These data support the use of RNase R prior to NGS analysis to increase the number of gene reads without increasing sequencing depth.
More Information
Applications
- Enrichment of circRNA prior to NGS or PCR analysis
- Removal of linear RNA from circRNA synthesis samples
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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