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% acrylamide 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.

Interactive Tools

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. Further information regarding NEB product quality can be found here.
  • 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.