RNase R HYJ412
RNase R (Ribonuclease R) is a 3'-5' ribonuclease from the Escherichia coli RNR superfamily that cleaves RNA into dinucleotides and trinucleotides in a stepwise manner from the 3' to the 5' direction. RNase R digests nearly all linear RNA molecules but is less sensitive to circular RNA, lariat structures, or double-stranded RNA with 3' overhangs of less than 7 nucleotides. RNase R is commonly used in gene expression and alternative splicing studies. It digests linear RNA to enrich for circRNAs and lariat RNAs. RNase R cleaves linear RNA to enrich for circRNAs, which are used to construct intronic cDNA libraries from total RNA.
Components
|
Components |
0.5 KU |
5 KU |
|
RNase R (20 U/μL) |
25 uL |
250 μL |
|
10×Reaction Buffer |
1 mL |
10×1 mL |
Product Information
|
Product Properties |
Clear liquid |
|
Source |
E.coli |
|
Activity Definition |
One unit of activity (U) is defined as the amount of enzyme required to convert 1 μg of poly-r(A) into acid-soluble nucleotides in a standard reaction at 37°C within 10 minutes. |
|
Optimum Reaction Temperature |
37℃ |
|
Storage Buffer |
50 mM Tris-HCl (pH 7.5 @ 25°C), 100 mM NaCl, 1mM DTT, 0.1mM EDTA, 50% Glycerol, 0.1% Triton® X-100 |
|
10×Reaction Buffer |
200 mM Tris-HCl (pH8.20 @25℃)，1 mM MgCl2，1 M KCl |
|
Storage Conditions |
-25 ~ -18℃, avoid repeated freezing and thawing |
Quality Control
1. Purity (SDS-PAGE): ≥95%.
2. Residual E.coli DNA: RT-qPCR assay using 10 μL of RNase R for 16S rDNA with a Ct value of ≥35.
3. Endonuclease Activity: No degradation was detected when 1 μg of dsRNA was incubated at 37°C for 4 hours in a 20 μL reaction containing at least 10 U of RNase R.
4. DNase Activity: No degradation was detected when 1 μg of pUC19 DNA was incubated at 37°C for 4 hours in a 10 μL reaction containing at least 10 U of RNase R.
5. Functional Assay: RNase R functionality was tested in a reaction containing 10 U of RNase R and 3 μg of total RNA from 293T cells. RNA abundance was assessed by RT-qPCR. The ΔCt values of linear RNAs were ≥4.5, and those of circular RNAs were ≤1, calculated with and without RNase R digestion.