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  • RNase If

    Description

    Ribonuclease If (RNase If) is an RNA endonuclease which will cleave at all RNA dinucleotide bonds leaving a 5´ hydroxyl and 2´, 3´ cyclic monophosphate (1). It has a preference for single-stranded RNA over double-stranded RNA. RNase If is a recombinant protein fusion of RNase I (from E. coli) and maltose-binding protein. It has identical activity to RNase I.

    Highlights

    • Isolated from a recombinant source
    • Eliminates single-stranded RNA from DNA and protein preparations
    • Supplied with 10X Reaction Buffer

    Product Source

    An E. coli strain containing a genetic fusion of the E. coli RNase I gene (2nd) and the gene coding for maltose binding protein (MBP).

    Reagents Supplied

    The following reagents are supplied with this product:

    Store at (°C)Concentration
    NEBuffer 3-2010X

    Advantages and Features

    Applications

    • Degradation of single-stranded RNA to mono-, di- and trinucleotide (3)
    • Used in ribonuclease protection assays

    Properties and Usage

    Unit Definition

    One unit is defined as the amount of enzyme required to fully digest 1 picomole of synthetic ssRNA 33-mer in a total reaction volume of 10 µl in 15 minutes in 1X NEBuffer 3 as visualized on a 20% acrylamine gel.

    Reaction Conditions

    1X NEBuffer 3
    Incubate at 37°C

    1X NEBuffer 3:
    100 mM NaCl
    50 mM Tris-HCl
    10 mM MgCl2
    1 mM DTT
    pH 7.9 @ 25°C

    Storage Temperature

    -20°C

    Storage Conditions

    10 mM Tris-HCl
    100 mM NaCl
    1 mM DTT
    0.5 mM EDTA
    50% Glycerol
    pH 8.0 @ 25°C

    Heat Inactivation

    70°C for 20 min

    Notes

    1. RNase If will not degrade DNA. It has a preference for single-stranded RNA over double-stranded RNA.

    References

    1. Spahr, P.F. and Hollingworth, B.R. (1961). J. Biol. Chem.. 236, 823-831.
    2. Meador, J. III and Kennell, D. (1990). Gene. 95, 1-7.
    3. Meador, J. III, Cannon, B., Cannistraro, V.J. and Kennell, D. (1990). Eur. J. Biochem.. 187, 549-553.

    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 info@neb.com or fill out the Technical Support Form for appropriate document.

    Quality Control

    Quality Assurance Statement

    • Purified free of endonucleases and contaminating exonucleases.

    Quality Control Assays

    The following Quality Control Tests are performed on each new lot and meet the specifications designated for the product. Individual lot data can be found on the Product Summary Sheet/Datacard or Manual which can be found in the Supporting Documents section of this page.
    • Endonuclease Activity (Nicking):
      The product is tested in a reaction containing a supercoiled DNA substrate. After incubation for 4 hours the percent converted to the nicked form is determined by agarose gel electrophoresis.
    • Exonuclease Activity (Radioactivity Release):
      The product is tested in a reaction containing a radiolabeled mixture of single and double-stranded DNA. After incubation for 4 hours the exonuclease activity is determined by the % release of radioactive nucleotides.
    • Single-Stranded DNase Activity (Radioactivity Release):
      The product is tested in reaction containing radiolabeled single-stranded DNA. After incubation the exonuclease activity is determined by the % release of radioactive nucleotides.

    Safety Data Sheet

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

    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 info@neb.com or fill out the Technical Support Form for appropriate document.