Product Class: Other

ShortCut® RNase III

icon_recomb icon_NEBU icon_incTemp37 icon_HI_no

Product Introduction

ShortCut RNase III converts long double-stranded RNA into a heterogeneous mix of short (18–25 bp) interfering RNAs (siRNA).

  • Isolated from a recombinant source
  • Useful for gene silencing and target validation
  • Supplied with 10X Reaction Buffer
Catalog # Size Concentration
M0245L 1,000 units 2,000 units/ml
M0245S 200 units 2,000 units/ml

Product Information

Description

ShortCut® RNase III, used with its manganese-containing reaction buffer, converts long double-stranded RNA into a heterogeneous mix of short (18–25 bp) interfering RNAs (siRNA) suitable for RNA interference in mammalian cells (1–3). 1.5 units (1 µl) of ShortCut RNase III is sufficient to convert 1 µg of dsRNA into siRNA suitable for RNA interference in mammalian cells.

ShortCut RNase III digestion of dsRNA: (A) Varying amounts of ShortCut RNase III were incubated with 2 µg of a 500 bp dsRNA for 20 minutes at 37°C in a 50 µl reaction. Digests were analyzed by 20% TBE polyacrylamide electrophoresis. Marker lane contains a mixture of 21 bp siRNA Marker and 100 bp DNA Ladder (NEB #N3231). (B) dsRNA fragments (1 kb and 175 bp) were digested with ShortCut RNase III. Digests were analyzed by 20% TBE polyacrylamide gel electrophoresis.
GFP Silencing in COS-7 Cells: COS-7 cells co-transfected in a 24 well plate with a plasmid expressing GFP in the absence (control) or the presence of 30 ng (4 nM) of GFP siRNA prepared using the ShortCut RNAi Kit. Cells were photographed 48 hours post-transfection.

Product Source

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

Reagents Supplied

The following reagents are supplied with this product:

Store at (°C) Concentration
Shortcut Reaction Buffer 10 X
10X EDTA -20 500 mM
MnCl2 -20 10 X
Glycogen RNase-Free 10 mg/ml
Product Categories:
miRNA & siRNA Products,
RNases Products
Applications:
RNA Modification

Advantages and Features

Application Features

  • Gene silencing
  • Target validation

Properties and Usage

Unit Definition

One unit is the amount of enzyme required to digest 1 µg of dsRNA to siRNA in 20 minutes at 37°C in a total reaction volume of 50 µl.

Reaction Conditions

1X ShortCut Reaction Buffer
Incubate at 37°C

Usage Concentration

1 - X

Storage Temperature

-20°C

Storage Conditions

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

Heat Inactivation

No

Unit Assay Conditions

One unit is the amount of enzyme required to digest 1 μg of dsRNA to siRNA in 20 minutes at 37°C in a total reaction volume of 50 μl.

References

  1. Morlighem, J.E. et al. (2007). Biotechniques. 42, 599-606.
  2. Yang, D. et al. (2002). Proc. Natl. Acad. Sci. USA. 99, 9942-9947.
  3. Calegari, F. et al. (202). Proc. Natl. Acad. Sci. USA. 99, 14236-14240.
  4. Donze, O. and Picard, D. (2002). Nucleic Acids Res. 30, e46.

Faqs & Tech Tips

FAQs

  1. What products does NEB offer for functional genomics and reverse genetics?

Protocols & Manuals

Protocols

  1. ShortCut RNase III Digestion Protocol (M0245)
  2. Purification of siRNA by Ethanol Precipitation Protocol (M0245)
  3. Transfection Guidelines for ShortCut® RNase III (M0245)

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.)

Other Tools & Resources

Interactive Tools

Quality & Safety

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 control's 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.