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Invitrogen™ RNaseOUT™ Recombinant Ribonuclease Inhibitor
Description
RNaseOUT Recombinant Ribonuclease Inhibitor is a potent noncompetitive inhibitor of pancreatic-type ribonucleases such as RNase A and is used to avoid RNA degradation in a variety of applications. It is an acidic protein with a molecular weight of ∼52 kDa that inhibits RNase A, RNase B, and RNase C.
The robust molecular structure of the RNaseOUT™ inhibitor together with a very high binding affinity for pancreatic-type RNases ensures superior protection of RNA from degradation making RNaseOUT™ highly effective and reliable in various molecular biology applications.
Features
- Stable under prolonged reaction conditions—effective RNA protection from RNases at:
- 4°C for at least 96 hours
- 25°C for at least 96 hours
- 37°C for 24 hours
- 50°C for up to 6 hours
- Oxidation resistant—highly stable and requires low concentrations of DTT to retain RNase inhibiting activity
- Wide activity temperature range—maintains full activity from 4°C to 55°C
- Superior quality—stringent quality control and industry leading manufacturing process
Applications
- In vitro transcription and translation
- CRISPR guide RNA production
- RNA purification
- cDNA synthesis
- RT-PCR
- RT-LAMP
- RNA-seq
- RNA modification and labeling
Source
Purified by affinity chromatography from E. coli expressing a cloned porcine gene.
Performance and quality testing
- SDS-PAGE purity
- Endo-deoxyribonuclease assay
- Protein concentration
- Specific activity
- Performance evaluated by RT-PCR
Unit definition
One unit inhibits 5 ng of RNase A by 50% using cytidine 2', 3' cyclic monophosphate (cCMP) as the substrate.
Unit reaction conditions
100 mM Tris-acetate (pH 6.5), 1 mM EDTA, 0.2 mM cCMP, 2 μg RNase A in 1 mL from 0 to 10 min at 25°C
Specifications
Specifications
| Concentration | 40 U/μL |
| Content And Storage | Store at -20°C. Avoid exposing product to frequent changes in temperature. RNaseOUT™ Ribonuclease Inhibitor requires 1 mM DTT to maintain activity. The product is guaranteed for 6 months from date of purchase unless otherwise stated in the product documentation. |
| Product Line | RNaseOUT |
| Quantity | 5000 Units |
Frequently Asked Questions (FAQs)
The following components are available as stand-alone items:
- Superscript III Reverse Transcriptase (Cat. Nos. 18080093, 18080044, 18080085)
- Oligo (dT)20 Primer (Cat. No. 18418020)
- Random hexamers (Cat. No. 48190011)
- 10 mM dNTP Mix (Cat. Nos. 18427013, 18427088)
- RNAseOUT Recombinant Ribonuclease Inhibitor (Cat. No. 10777019)
- E. coli RNAse H (Cat. Nos. 18021014, 18021071)
The inhibitor acts as a safeguard against degradation of target RNA due to ribonuclease contamination of the RNA preparation.
RNaseOUT Recombinant Ribonuclease Inhibitor is provided at a concentration of 40 U/µL.
RNaseOUT Recombinant Ribonuclease Inhibitor (Cat. No. 10777019) unit reaction conditions are defined at 25 degrees C. The upper temperature limit for full functionality is 40 degrees C. The enzyme half-life is decreased as temperatures increase above 40 degrees C. There is some residual activity up to 50-55 degrees C but heating at 65 degrees C will inactivate the enzyme. As RNaseOUT can be used in various applications like cDNA synthesis, RT-PCR, and in vitro transcription, the recommendation is to follow the temperature settings required for the respective method.
RNAlater should not impact downstream PCR amplification as long as the sample has been properly cleaned before proceeding with DNA isolation, as stated in the RNAlater Stabilization Solution manual (https://assets.thermofisher.com/TFS-Assets/LSG/manuals/7020M.pdf) on page 7.
For Research Use Only. Not for use in diagnostic procedures.