To access your account, log in or register.	shopping cart | log in

FAQs for RNAi and RNA Enzymes Technical Reference for RNAi and RNA Enzymes Enzyme Finder NEBcutter NEBuffer Chart Double Digest Finder Isoschizomers DNA Sequences and Maps REBASE 6-Tube Magnetic Separation Rack Biotinylated T7 Primer (25-mer) HiScribe RNAi Transcription Kit LITMUS 28i Vector LITMUS 38i Vector ProtoScript® First Strand cDNA Synthesis Kit ShortCut® RNAi Kit siRNA Marker Streptavidin Magnetic Beads

Home > Products > RNAi and RNA Enzymes > RNases > ShortCut® RNase III ShortCut® RNase III Catalog # Size Concentration Price Qty   M0245L 650 units 1,300 units/ml $420.00 M0245S 130 units 1,300 units/ml $105.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF Isolated from a recombinant source Gene silencing Target validation Supplied with 10X Reaction Buffer 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 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. 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). RNase III is cleaved from the fusion and purified away from MBP. Applications: Gene silencing Target validation Reagents Supplied: EDTA (250 mM) MnCl2 (200 mM) RNase-free Glycogen (10 mg/ml) ShortCut Reaction Buffer Enzyme Properties Heat Inactivation: No Reaction & Storage Conditions Reaction Conditions: 1X ShortCut Reaction Buffer Supplemented with 10 mM MnCl2 Incubate at 37°C. 1X ShortCut Reaction Buffer: 50 mM Tris-HCl 1 mM Dithiothreitol pH 7.5 @ 25°C 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. Concentration: 1,300 units/ml Storage Conditions: 10 mM Tris-HCl 500 mM NaCl 1 mM Dithiothreitol 0.5 mM EDTA 50% Glycerol pH 8.0 @ 25°C Storage Temperature: -20°C 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. References Yang, D. et al (2002) Proc. Natl. Acad. Sci. USA, 99, 9942-9947. Calegari, F. et al. (1988) Proc. Natl. Acad. Sci. USA, 99, 14236-14240. Donze, O. and Picard, D. (2002) Nucl. Acids Res., 30, e46. Companion Products 6-Tube Magnetic Separation Rack Biotinylated T7 Primer (25-mer) HiScribe RNAi Transcription Kit LITMUS 28i Vector LITMUS 38i Vector ProtoScript® First Strand cDNA Synthesis Kit ShortCut® RNAi Kit siRNA Marker Streptavidin Magnetic Beads Legal Licenses/Patents/Disclaimers: Notice to Buyer/User: Commercial use of ShortCut® RNAse III may require a license from New England Biolabs, Inc. Use of ShortCut® RNAse III for RNA interference may require a license from the Carnegie Institution of Washington. For further information, contact the Director of Administration and Finance at the Carnegie Institute of Washington at 1530 P Street N.W., Washington, DC 20005–1910. Tel: 202-939-1118.