Product Class: Other

RNase HII
NEBU recombinant 37 No

Product Introduction

  • DNA endonuclease
  • Catalyzes the cleavage of the DNA phosphodiester backbone 5' to ribonucleotide, or string of ribonucleotides, embedded within dsDNA leaving a 3' OH and 5' phosphate
  • Not sure which RNase to choose? Learn more about the different RNases offered by NEB.
Catalog # Size Concentration
M0288S 250.0 units 5000 units/ml
M0288L 1250.0 units 5000 units/ml

Product Information

Description

Ribonuclease HII (RNase HII) is an endoribonuclease that preferentially nicks 5´ to a ribonucleotide within the context of a DNA duplex. The enzyme leaves 5´ phosphate and 3´ hydroxyl ends (1). RNase HII will also nick at multiple sites along the RNA portion of an Okazaki fragment.

Product Source

An E. coli strain containing a genetic fusion of the RNase HII gene (rnhB) from E. coli and the gene coding for maltose binding protein (MBP). Following affinity chromatography, RNase HII is cleaved from the fusion construct by Factor Xa and then purified away from both MBP and Factor Xa. RNase HII cleaved from MBP has four additional amino acids at its N-terminus (Ile-Ser-Glu-Phe).
This product is related to the following categories:
RNases,
Epitranscriptome Analysis,

Reagents Supplied

Reagents Supplied

The following reagents are supplied with this product:

NEB # Component Name Component # Stored at (°C) Amount Concentration

Properties & Usage

Unit Definition

One unit is defined as the amount of enzyme required to yield a fluorescence signal consistent with the nicking of 100 picomol of synthetic double-stranded DNA substrate containing a single ribonucleotide near the quencher of a fluorophore/quencher pair in 30 minutes at 37°C in 1X ThermoPol Buffer.

Reaction Conditions

1X ThermoPol® Reaction Buffer
Incubate at 37°C

1X ThermoPol® Reaction Buffer
20 mM Tris-HCl
10 mM (NH4)2SO4
10 mM KCl
2 mM MgSO4
0.1% Triton® X-100
(pH 8.8 @ 25°C)

Storage Buffer

20 mM Tris-HCl
100 mM NaCl
1 mM DTT
1 mM EDTA
50% Glycerol

Heat Inactivation

No

Unit Assay Conditions

1X ThermoPol Reaction Buffer with 30 nM of a synthetic dsDNA 26-mer containing an internal ribonucleotide in a total reaction volume of 150 µl.

Application Features

Nicking of products generated with a polymerase that will incorporate ribonucleotides
Generation of a double-stranded break at the site of an incorporated ribonucleotide when used with T7 Endo I
Degradation of the RNA portion of Okazaki fragments

Product Notes

  1. RNase HII displays reduced hybridase activity compared to RNase H (2).
  2. Okazaki fragments are preferentially nicked 5´ to the ribonucleotide at the junction of the RNA/DNA sequence (1).
  3. RNase HII prefers a dsDNA duplex containing a single ribonucleotide over a RNA/DNA hybrid substrate (1) or an Okazaki fragment (3).
  4. Incubation with 0.1% SDS is sufficient to inactivate RNase HII.

References

  1. Rydberg, B. and Game, J. (2002). Proc. Natl. Acad. Sci.. 99,
  2. Itaya, M. (1990). Proc. Natl. Acad. Sci.. 87,
  3. Nichols, N.M. Unpublished Observation.

Protocols, Manuals & Usage

Protocols

  1. Nicking a single ribonucleotide site within a dsDNA substrate or an Okazaki fragment with RNase HII (NEB #M0288)

Tools & Resources

Selection Charts

FAQs & Troubleshooting

FAQs

  1. RNase HII cannot be heat-inactivated. How can RNase HII be inactivated?
  2. Which side of the ribonucleotide does RNase HII cut?

Quality, Safety & Legal

Quality Assurance Statement

Quality Control tests are performed on each new lot of NEB product to meet the specifications designated for it. Specifications and individual lot data from the tests that are performed for this particular product can be found and downloaded on the Product Specification Sheet, Certificate of Analysis, data card or product manual. Further information regarding NEB product quality can be found here.

Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Certificate Of Analysis

The Certificate of Analysis (COA) is a signed document that includes the storage temperature, expiration date and quality controls for an individual lot. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]_[Lot Number]

Safety DataSheets

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Legal and Disclaimers

Products and content are covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB). The use of trademark symbols does not necessarily indicate that the name is trademarked in the country where it is being read; it indicates where the content was originally developed. The use of this product may require the buyer to obtain additional third-party intellectual property rights for certain applications. For more information, please email busdev@neb.com.

This product is intended for research purposes only. This product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

New England Biolabs (NEB) is committed to practicing ethical science – we believe it is our job as researchers to ask the important questions that when answered will help preserve our quality of life and the world that we live in. However, this research should always be done in safe and ethical manner. Learn more.

Trademarks

NEW ENGLAND BIOLABS® is a registered trademark of New England Biolabs, Inc. 

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