• For digestion of full RNA, longer incubations of 2-Three hours are generally required.
• If degradation is inefficient, use a barely elevated incubation temperature (40-45°C) and complement additional enzyme partway (e.g. 0.5 μl after 1 hour) by way of the method. The higher temperature is particularly useful for degrading extraordinarily structured linear RNAs, equal to rRNAs. Do not exceed 45°C or incubate over 3
hours, as this may end in non-enzymatic RNA degradation.
• RNase R shows low train on tRNA, rRNA and totally different extraordinarily structured RNAs, for which the three’ end is double stranded with a short 3’ overhang. These RNA species can stall the enzyme and finish in drastically lowered train. If inefficient degradation is seen, it is endorsed to each upscale the digestion, use additional RNase R,
or take away rRNA from full RNA extracts earlier to digestion.
• Keep in mind that spherical RNAs symbolize a small proportion of full RNA (generally 0.1%-0.01%), because of this reality RNase R treatment will most actually finish in low ranges of RNA (picogram-range), presumably undetectable by most methods. For that purpose, a starting amount of not lower than 10 µg of full RNA is useful for a lot of downstream features.
• Whereas the enzyme could also be heat inactivated the method should not be useful since extreme heat can lead to RNA hurt. Phenol-chloroform precipitation could be utilized instead. For NGS, robust half reversible immobilization (SPRI) bead cleanup is useful.
• Magnesium at concentrations of 0.1-1.Zero mM is required for optimum train. If ETA is
present, compensate by together with MgCl2 to 1.Zero Mm final focus.
DESCRIPTION:
RNase R is an E. coli exoribonuclease which shows 3’-to-5’ exonuclease train, successfully digesting virtually all linear RNA species. This enzyme does not digest spherical, lariat, or double stranded RNA with temporary 3’ overhangs (decrease than sevennucleotides). As such, this enzyme is ideally suited to the study of lariat RNA produced by typical splicing, along with circRNAs which come up by way of back-splicing.
APPLICATIONS:
• Enriching circRNAs in natural samples
• Identification of intronic lariat sequences
• Identification of exonic circRNAs
• Studying varied splicing
• Manufacturing of artificial spherical RNAs
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.
Stay away from repeated freeze-thaw cycles of all components to retain most effectivity. All components
are regular for one 12 months from the date of transport when saved and handled appropriately.
Cat # +Dimension
M1228-500
Dimension
500 U
Highlights
RNase R is an E. coli exoribonuclease which shows 3’-to-5’ exonuclease train, successfully digesting virtually all linear RNA species.
Storage Conditions
-20°C
Supply Conditions
Gel Pack
USAGE
For Evaluation Use Solely! Not For Use in Individuals.
Description: RNase L is an enzyme that in humans is encoded by the RNASEL gene. This gene encodes a component of the interferon-regulated 2-5A system that functions in the antiviral and antiproliferative roles of interferons. Mutations in this gene have been associated with predisposition to prostate cancer and this gene is a candidate for the hereditary prostate cancer 1 (HPC1) allele.
Description: RNAse H2A Antibody: Ribonucleases (RNAses) H are enzymes that hydrolyze the RNA strands of RNA/DNA hybrids. The major role of these enzymes is to remove the RNA strand from the RNA/DNA hybrids that form during DNA replication and repair. RNAse H2 is made up of three subunits; all three are required for RNAse activity. Recent evidence has demonstrated that mutations in RNAse H2A or any of the other subunits result in Aicardi-Goutieres syndrome (AGS), a neurological disorder with similar symptoms to viral brain infections including high levels of IFN-alpha in the cerebral spinal fluid. Similar conditions are observed with mutations in TREX1, a single-stranded DNA exonuclease, suggesting that RNAse H2 and TREX1 may have similar roles, and that mutations in any of these genes lead to an accumulation of intracellular nucleic acids, triggering an inflammatory response through activation of the innate immune system.
Description: RNAse H2A Antibody: Ribonucleases (RNAses) H are enzymes that hydrolyze the RNA strands of RNA/DNA hybrids. The major role of these enzymes is to remove the RNA strand from the RNA/DNA hybrids that form during DNA replication and repair. RNAse H2 is made up of three subunits; all three are required for RNAse activity. Recent evidence has demonstrated that mutations in RNAse H2A or any of the other subunits result in Aicardi-Goutieres syndrome (AGS), a neurological disorder with similar symptoms to viral brain infections including high levels of IFN-alpha in the cerebral spinal fluid. Similar conditions are observed with mutations in TREX1, a single-stranded DNA exonuclease, suggesting that RNAse H2 and TREX1 may have similar roles, and that mutations in any of these genes lead to an accumulation of intracellular nucleic acids, triggering an inflammatory response through activation of the innate immune system.
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