RNase R is an E. coli

APPLICATIONS:

• Enriching circRNAs in pure samples
• Identification of intronic lariat sequences
• Identification of exonic circRNAs
• Discovering out fairly a number of splicing
• Manufacturing of artificial spherical RNAs

DESCRIPTION:

RNase R is an E. coli exoribonuclease which displays 3’-to-5’ exonuclease observe, effectively digesting almost all linear RNA species. This enzyme does not digest spherical, lariat, or double stranded RNA with non everlasting 3’ overhangs (decrease than sevennucleotides). As such, this enzyme is ideally suited to the take a look at of lariat RNA produced by typical splicing, along with circRNAs which come up by means of back-splicing. By eradicating linear
RNAs from cellular or RNA extracts, RNase R drastically facilitates the identification of spherical species by means of RNA-sequencing. This allows researchers to probe the panorama of splicing events with higher depth.

 

EZYME UNIT DEFINITION: One unit is printed as the amount 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.

Avoid repeated freeze-thaw cycles of all components to retain most effectivity. All components
are frequent for one 12 months from the date of transport when saved and handled exactly.

Cat # +Dimension	M1228-500
Dimension	500 U
Highlights	RNase R is an E. coli exoribonuclease which displays 3’-to-5’ exonuclease observe, effectively digesting almost all linear RNA species.
Storage Conditions	-20°C
Present Conditions	Gel Pack
USAGE	For Evaluation Use Solely! Not For Use in Individuals.

SAMPLE PROCESSING GUIDELINES AND TROUBLESHOOTING:

• For digestion of full RNA, longer incubations of 2-Three hours are sometimes required.
• If degradation is inefficient, use a barely elevated incubation temperature (40-45°C) and complement further enzyme partway (e.g. 0.5 μl after 1 hour) by means of the tactic. The higher temperature may very well be very useful for degrading terribly structured linear RNAs, equal to rRNAs. Do not exceed 45°C or incubate over 3
hours, because it ought to end in non-enzymatic RNA degradation.

• RNase R displays low observe on tRNA, rRNA and utterly utterly completely different terribly structured RNAs, for which the three’ end is double stranded with a quick 3’ overhang. These RNA species can stall the enzyme and finish in drastically lowered observe. If inefficient degradation is observed, it is endorsed to each upscale the digestion, use further RNase R,
or take away rRNA from full RNA extracts earlier to digestion.

• Perceive that spherical RNAs symbolize a small proportion of full RNA (sometimes 0.1%-0.01%), as a consequence of this actuality RNase R remedy will most positively finish in low ranges of RNA (picogram-range), presumably undetectable by most methods. For that motive, a starting amount of not lower than 10 µg of full RNA is useful for plenty of downstream capabilities.

• Whereas the enzyme could also be heat inactivated the tactic should not be useful since extreme heat may end up in RNA hurt. Phenol-chloroform precipitation is also utilized as a alternative. For NGS, sturdy half reversible immobilization (SPRI) bead cleanup is useful.

• Magnesium at concentrations of 0.1-1.Zero mM is required for optimum observe. If ETA is
present, compensate by together with MgCl2 to 1.Zero Mm final focus.

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RNase R
M1228-500	Biovision		EUR 365

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