RNase I

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Reagents Supplied

10X RNase I Reaction Buffer

Description

• Only available RNase that cleaves the phosphodiester bond of all four bases
• Prefers single-stranded RNA to double-stranded RNA
• Produced from an overexpressing clone in E. coli (2)
• Contains no endonuclease or exonuclease activity toward DNA substrates
• No need for boiling prior to use
• Pure, contains no endonuclease or exonuclease activity toward DNA substrates

Applications

• Degrades RNA to cyclic nucleotide monophosphates leaving a 5'-OH and 2'-, 3'-cyclic monophosphate
• Ideal for ribonuclease protection assays
• Useful for mapping or quantitation of RNA by selective cleavage of single-strand regions

Unit Definition

One unit is the amount of enzymes required to degrade 50% of [α-³³P] labeled RNA transcript mixed with 2 µg of yeast RNA in 30 minutes at 37°C as detected by TCA precipitation

Storage Buffer

10 mM Tris-HCl (pH 8.0 at 22°C)
200 mM NaCl
50% glycerol

Quality Control

All preparations are assayed for contaminating exonuclease and nonspecific endonuclease activities

Storage Conditions

Store at -20°C
Product shipped on Dry Ice

References

(1) Meador, J. III, and Kennell, D. (1990) Gene 95, 1-7
(2) Meador, J. et al. (1990) Eur. J. Biochem. 187, 549