3rd July 2024

RNase III, E. coli

Applications

  • Digestion of long dsRNA to short dsRNA.
  • RNA structure studies.1
  • RNA processing and maturation studies.2-4

Ribonuclease III (RNase III) from E. coli is an endoribonuclease that specifically digests dsRNA to dsRNA fragments that have two-base, 3´ overhangs.5,6 Complete digestion of dsRNA results in dsRNA fragments of 12-15 bp.

RNase III can digest dsRNA composed of canonical ribonucleotides as well as DuraScript® RNA produced using Epicentre’s DuraScribe® T7 Transcription Kit, which contains 2´-fluorine-CMP and 2´-fluorine-UMP in place of the canonical CMP and UMP nucleotides.

Unit Definition: One unit of RNase III, E. coli, solubilizes 1 nmol of ribonucleotide in 30 minutes at 37°C using poly(A)-poly(U) as the substrate under standard assay conditions.

Quality Control: RNase III is tested to specifically digest dsRNA transcripts in a mixture containing dsRNA, ssRNA, and dsDNA.

Figure 1. RNase III rapidly digests long dsRNA into short dsRNA. Five micrograms of a 1.4-kb dsRNA were digested using 1 unit of RNase III. Reaction aliquots were removed after 1 minute and 2 minutes of digestion and analyzed by 12% PAGE. Lane 1, 1.4-kb dsRNA; Lane 2, 1.4-kb dsRNA after 1 minute digestion with RNase III; Lane 3, 1.4-kb dsRNA after 2 minute digestion with RNase III.Storage Buffer: 50% glycerol containing 50 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT, 0.5 M NaCl, and 0.1% Triton® X-100.

References

  1. Evguenieva-Hackenberg, E. and Klug, G. (2000) J. Bacteriol. 182, 4719.
  2. Nicholson, A. (1999) FEMS Microbiol. Rev. 23, 371.
  3. Grunberg-Manago, M. (1999) Annual Rev. Genet. 33, 193.
  4. Drider, D. et al. (1999) J. Mol. Microbiol. Biotechnol. 1, 337.
  5. Robertson, H.D. et al. (1968) J. Biol. Chem. 243, 82.
  6. Lamontagne, B. et al. (2001) Curr. Issues Mol. Biol. 3, 71.

Ordering

Catalog No. Concentration Size
RNase III, E. coli
RN02950 1 U/µl 50 Units