RNase I (DNase free)
recombinant, E. coli
For in vitro use only!
Definição de unidade: One unit is the amount of enzyme required to degrade 1 μg of RNA in 30 minutes at 37 °C.
Envio: shipped on blue ice
Condições de armazenamento: store at -20 °C
avoid freeze/thaw cycles
Validade: 12 months
Peso molecular: 27.0 kDa
Pureza: > 95 % (SDS-PAGE)
Forma: liquid (Supplied in 10 mM Tris-HCl pH 8.0, 200 mM NaCl and 50 % [v/v] glycerol)
- degrades RNA to cyclic nucleotide monophosphates leaving a 5'-OH and 2'-, 3'-cyclic monophosphate
- Ribonuclease Protection Assays
- Mapping or Quantitation of RNA by selective cleavage of single-strand regions
- Eliminates RNA from DNA- and protein purification
Ribonuclease l (27 kD) is a completely nonspecific ribonuclease that hydrolyzes the phosphodiester bond of all four bases.
It degrades any RNA to a mixture of mono-, di-, and trinucleotides and does not degrade DNA, although it does bind to DNA. It has a marked preference for single-stranded RNA over double-stranded RNA, which allows it to work well in RNase Protection Assays. It has a high specific activity which, coupled with its non-specificity, typically results in complete degradation of RNA using ng amounts of protein.
Contains no endonuclease or exonuclease activity toward DNA substrates.
For RNase Protection Assays using approximately 10 μg of total RNA per sample, we suggest using 100 - 500 units of enzyme at 37 °C for 30 min.
For boiling lysate minipreps we suggest using 50 units at 37 °C for 30 min.
- does not require divalent cations and is fully active in Tris and phosphate buffers
- 80 % active in 0.3 M NaCl and 100 % active in 0.1 - 0.2 M NaCl
- completely and irreversibly inactivated by 0.1 % SDS or phenol extraction
- inactivated by freeze-thaw cycles in aqueous buffers but is protected from inactivation by 20 % glycerol
Meador et al. (1990) Cloning and sequencing the gene encoding Escherichia coli ribonuclease I: exact physical mapping using the genome library. Gene 95: 1.
Ono et al. (1987) Nucleotide sequence of the pnd gene in plasmid R483 and role of the pnd gene product in plasmolysis. Mirobiol. Immunol.31:1071.
Ito et al. (1983) The roles of RNA polymerase and RNAase I in stable RNA degradation in Escherichia coli carrying the srnB+gene. Biochim. Biophys. Acta 739: 27.