Description:
Description
Ribonuclease A (RNase A) is a highly purified ribonuclease expressed in E.coli and purified by genetic engineering technology, with a molecular weight of 13.7 kDa and four pairs of disulfide bonds. The enzyme is an endonuclease that can specifically recognize and hydrolyze the phosphate diester bond formed by the phosphate group on 5'-ribose of nucleotide and 3'-ribose of adjacent pyrimidine nucleotides. This product has the best activity of hydrolyzing single strand RNA, and has activity under various reaction conditions. At low salt concentration (0-100 mM NaCl), it can be used to cut RNA strands in single stranded RNA, double stranded RNA and RNA/DNA hybrid chains. While at high salt concentration (≥ 0.3M), RNase A only hydrolyzes single stranded RNA. This enzyme is often used to remove RNA during the preparation of plasmid DNA or genomic DNA. This product does not contain DNase and protease and can be used directly. In addition, the product can also be used in molecular biology experiments such as RNA enzyme protection analysis and RNA sequence analysis.
Appearance
White or pain white lyophilized powder
Package
100mg/Vial
Stocking buffer
This product is mainly used to hydrolyze and remove RNA during DNA preparation:
(1) Micro-extraction of plasmid: RNase A was added to the restriction enzyme digestion reaction solution, and the final concentration was 20 μg/mL。
(2) Large scale extraction of plasmid: RNase A is added to the cell lysate, with the final concentration of 20 μg/mL, 37℃, 20~60 min, then extract with phenol chloroform, and finally precipitate with 2M ammonium acetate and twice the volume of 100% ethanol.
[Note]: The addition of EDTA can inhibit the possible DNase activity.
Unite definition
At 25℃, 100 mM Tris acetate (pH 6.5), 1 mM EDTA and 1 mM cyclic 2', 3'-CMP reaction solution, the amount of enzyme that increases A286 per milliliter per minute by 0.1 is defined as an enzyme activity unit (Kunitz unit).
Stability
Stable at -20℃for 36 months.
