RNase R

Product Description:

RNase R is a 3 '-5' exonuclease derived from the Escherichia coli RNR superfamily that progressively cuts RNA into dinucleotides and trinucleotides from the 3 '-5' direction. RNase R can digest almost all linear RNA molecules, but it is difficult to digest circular RNA, lasso structures, or double-stranded RNA molecules with fewer than 7 nucleotides at the 3 'protruding end.

RNase R is an essential tool enzyme for circRNA identification and enrichment experiments, digesting linear RNA to enrich circRNA or lariat RNA.

Fig.1: Schematic diagram of RNase R digested RNA.

Application

1. Identification of circRNA: To prove whether the molecule is a circRNAbased on whether bands were detected in the RNase R(+) and RNase R(-) groups.

Fig.2: RT-PCR detection of Lariatt/Circular RNA after RNase R digestion（Suzuki H et al., 2006）

2. Enrichment of circRNA: High-throughput sequencing often requires enrichment of circRNA. In order to explore the enrichment degree of circRNA and the changes of related genes after digestion of RNase R, the samples of RNase R(+) and RNase R(-) groups can be sequenced simultaneously.

Fig.3: RNase R(+) and RNase R(-) RNA sequencing diagram (Jeck WR et al., 2014)

3. Purification of circRNA: Eliminating the interference of linear RNA residues is the key to artificial synthesis of high-purity circRNA. Efficient RNase R is not only a key tool for ligase circRNA synthesis, but also essential in the intron self-splicing circRNA artificial synthesis.

Product Features:

Strong specificity: specific digestion of linear RNA

Efficient and fast: most of the linear RNA can be digested in 5-15min

Buffer compatibility: Digestive products can be used directly in downstream experiments;

Easy to use: simple system, one-step reaction at 37℃

Performance comparison

1. RNA electrophoretic detection

Fig.4: Added 10U RNase R to 2.5 μg total RNA, incubated at 37℃ for 15 min, and then directly detected by electrophoresis. The results showed that the band of RNase R(+) group became fainter(invisible), indicating that RNase R had digestive effect on total RNA. RNase R from Geneseed and Company A or Company E is comparable for total RNA digestion.

2. RT-qPCR test

Fig.5: Added 10U RNase R to 2.5 μg total RNA, and RT-qPCR experiments were performed after incubation at 37℃ for 30 min. The results showed that the abundance of both β-actin and FGFR2 in the RNase R+ group was significantly reduced, suggesting that RNase R could digest the linear RNA. RNase R from Geneseed and Company E is comparable for linear RNA digestion, and outperforms Company A.

Fig.6: Added 10U RNase R was added to 2.5 μg total RNA, and RT-qPCR experiments were performed after incubation at 37℃ for 30 min. The results showed that the abundance of hsa_circFOXO3_002 and hsa_circMTO1_001 in the RNase R+ group was basically unchanged, indicating that circRNA was resistant to digestion of RNase R. The effect of Geneseed’s RNase R is comparable to that of Company E or Company A.

Note: The data were calculated using the "RNase R+ Geneseed" group as the control, and its relative abundance value was set as 1.

 Kit Contents:

Product information

Storage and Handling:

Store the kit at -20℃. The kit is stable for at least 18 months from date of receipt.