DESCRIPTION: RNase R is an E. coli exoribonuclease which displays 3’-to-5’ exonuclease exercise, effectively digesting practically all linear RNA species. This enzyme doesn’t digest round, lariat, or double stranded RNA with brief 3’ overhangs (lower than sevennucleotides). As such, this enzyme is ideally suited to the examine of lariat RNA produced by conventional splicing, in addition to circRNAs which come up by means of back-splicing. By eradicating linear
RNAs from mobile or RNA extracts, RNase R drastically facilitates the identification of round species by means of RNA-sequencing. This permits researchers to probe the panorama of splicing occasions with higher depth.
APPLICATIONS:
• Enriching circRNAs in organic samples
• Identification of intronic lariat sequences
• Identification of exonic circRNAs
• Learning various splicing
• Manufacturing of synthetic round RNAs
EZYME UNIT DEFINITION: One unit is outlined as the quantity of RNase R that converts 1
µg of poly(A) into acid soluble nucleotides in 10 minutes at 37°C. ENZYME STORAGE BUFFER: 50 mM Tris-HCl (pH 7.5), 100mM NaCl, 0.1 mM EDTA, 1 mM DTT, and 50% (v/v) Glycerol 10X RNase R REACTION BUFFER COMPONENTS: 200 mM Tris-HCl, 1 M KCl, 1 mM, MgCl2, pH 7.5.
STORAGE CONDITIONS: Retailer at -20°C.
Keep away from repeated freeze-thaw cycles of all parts to retain most efficiency. All parts
are steady for one 12 months from the date of transport when saved and dealt with correctly.
Cat # +Dimension
M1228-500
Dimension
500 U
Highlights
RNase R is an E. coli exoribonuclease which displays 3’-to-5’ exonuclease exercise, effectively digesting practically all linear RNA species.
Storage Situations
-20°C
Delivery Situations
Gel Pack
USAGE
For Analysis Use Solely! Not For Use in People.
Biovision rnase-r
SAMPLE PROCESSING GUIDELINES AND TROUBLESHOOTING:
• For digestion of complete RNA, longer incubations of 2-Three hours are sometimes required.
• If degradation is inefficient, use a barely increased incubation temperature (40-45°C) and complement extra enzyme partway (e.g. 0.5 μl after 1 hour) by means of the process. The upper temperature is especially helpful for degrading extremely structured linear RNAs, equivalent to rRNAs. Don’t exceed 45°C or incubate over 3
hours, as this will result in non-enzymatic RNA degradation.
• RNase R displays low exercise on tRNA, rRNA and different extremely structured RNAs, for which the three’ finish is double stranded with a brief 3’ overhang. These RNA species can stall the enzyme and end in drastically lowered exercise. If inefficient degradation is noticed, it’s endorsed to both upscale the digestion, use extra RNase R,
or take away rRNA from complete RNA extracts previous to digestion.
• Remember that round RNAs symbolize a small proportion of complete RNA (sometimes 0.1%-0.01%), due to this fact RNase R remedy will most certainly end in low ranges of RNA (picogram-range), presumably undetectable by most strategies. For that reason, a beginning quantity of not less than 10 µg of complete RNA is beneficial for many downstream functions.
• Whereas the enzyme may be warmth inactivated the process shouldn’t be beneficial since excessive warmth can result in RNA harm. Phenol-chloroform precipitation can be utilized as a substitute. For NGS, strong part reversible immobilization (SPRI) bead cleanup is beneficial.
• Magnesium at concentrations of 0.1-1.Zero mM is required for optimum exercise. If ETA is
current, compensate by including MgCl2 to 1.Zero Mm ultimate focus.
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