■ RNase H content
Ribonuclease H (20 to 60 U/μl) 600 U
5×Hybrid RNA Degeneration Buffer 300 μl
■ RNase H description
RNase H is an endonuclease that specifically breaks down RNA strands in RNA-DNA hybrids with a molecular weight of 21,000 and an optimal pH of about 8.0. The active factor is Mg2+ or Mn2+.
■ Preservation
-20℃.
■ Origin
Escherichia coli HB101 containing rnh plasmid (pKH11) and regulator plasmid (pNT203)
■ Activity definition
Using poly(rA)·poly(dT) as substrate, the amount of enzyme required to produce 1 nmol of acid soluble substance within 20 minutes at 30℃and pH7.7 was defined as 1 unit of activity (U).
■ Purity
After the reaction of 1.50 U of this enzyme and 1 μg of λ DNA-Hind III at 37℃for 16 h, the electrophoretic bands of DNA did not change.
When 2.50 U of this enzyme and 1 μg of superhelical pBR322 DNA were reacted at 37℃for 1 hour, the electrophoretic bands of DNA did not change.
When 3.50 U of this enzyme and 1 μg of 16S, 23S rRNA reacted at 37℃for 1 hour, the electrophoretic bands of RNA did not change.
■ Use
1. cDNA Cloning was conducted by Okayama-Berg method.
2. Detection of DNA-RNA hybrids.
3. Remove the Poly (A) end of mRNA in the presence of Oligo (dT).
■ Use attention
This enzyme is purified from E. coli recombinant with few impurities and is a product of high purity. Therefore, even the high concentration of products, a large number of use is not a problem.
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