RNase I (DNase free)
Endonuclease
recombinant, E. coli
Catálogo Nº | Apresentação | Preço (R$) | Comprar |
---|---|---|---|
EN-176S | 2.000 units | Sob demanda | Adicionar ao Carrinho |
EN-176L | 5 x 2000 units | Sob demanda | Adicionar ao Carrinho |
For general laboratory use.
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 gel packs
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)
Formulários:
- 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
Descrição:
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.
Reaction conditions:
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.
Attention:does not require divalent cations and is fully active in Tris and phosphate buffers80 % active in 0.3 M NaCl and 100 % active in 0.1 - 0.2 M NaClcompletely and irreversibly inactivated by 0.1 % SDS or phenol extractioninactivated by freeze-thaw cycles in aqueous buffers but is protected from inactivation by 20 % glycerol
Referências selecionadas:
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.