Product Class: Other

RNase HII
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Item# Description List Price Web Price Qty
M0288L RNase HII - 1,250 units $412.00
$370.80
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M0288S RNase HII - 250 units $102.00
$91.80
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*On-line ordering is for Canadian customers only. Web pricing is applicable only to orders placed online at www.neb.ca
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Companion Products

Item# Description List Price Web Price Qty
B9004S ThermoPol Reaction Buffer Pack - 6.0 ml $31.00
$27.90
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    Product Introduction

    Ribonuclease HII (RNase HII) is an endoribonuclease that preferentially nicks 5´ to a ribonucleotide within the context of a DNA duplex.

    • Isolated from a recombinant source
    • Supplied with 10X reaction buffer
    • Tested for the absence of DNases and RNases

    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).

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C) Concentration
    ThermoPol® Reaction Buffer Pack -20 10X
    Product Categories:
    RNases Products

    Advantages and Features

    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

    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 Pack
      Incubate at 37°C

      1X ThermoPol® Reaction Buffer Pack
      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 Temperature

      -20°C

      Storage Conditions

      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.

      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. Reaction Protocol for RNase HII (M0288)

      Datacards

      The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at [email protected] or fill out the Technical Support Form for appropriate document.)

      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?

      Troubleshooting

      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

      This product is covered by one or more patents, trademarks and/or copyrights owned or controlled by New England Biolabs, Inc (NEB).

      While NEB develops and validates its products for various applications, the use of this product may require the buyer to obtain additional third party intellectual property rights for certain applications.

      For more information about commercial rights, please contact NEB's Global Business Development team at [email protected].

      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.

      Trademarks

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

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