Ribonuclease H, E.coli, Recombinant (RNase H) (5 x 1000 units)
Product#: FNK-02-060-5
$330.28
Availability:
Ships in 1-2 Weeks
Ribonuclease H, E.coli, Recombinant
Store at -20°C
Size 1000 Units / 1000 Units x5
Order Information
| Cat No. | Size | Link |
| FNK-02-060 | 1000 units |  |
| FNK-02-060-5 | 5 x 1000 units | |
Description
Ribonuclease H (RNase H) is an endoribonuclease which specifically degrades the RNA strand of an RNA/DNA hybrid, leaving the DNA strand and unhybridized RNA intact. E.coli RNase H (=RNaseHI) was over-expressed in E. coli as a recombinant protein and highly purified. MW is 17.6 kDa.
Fig.1 SDS-PAGE of E.coli RNaseH
- Removal of mRNA in DNA/RNA hybrid prior to the synthesis of the second strand of cDNA (1, 2)
- Removal of poly (A) tails from mRNA after hybridization with oligo (dT) (3)
- Oligodeoxyribonucleotide-directed site-specific cleavage of RNA (4)
Form 50 units/ul in 20mM Tris-HCl (pH 7.5), 100mM KCl, 1mM DTT, 50% Glycerol
Specific Activity 100,000 units/mg protein
Unit Definition
1 unit is defined as the amount of the enzyme that hydrolyzes 1 nmol of the RNA in 3H labeled M13 DNA/RNA hybrid to acid-soluble ribonucleotides in 20 min at 37℃.
Quality Assurance
Greater than 95% protein determined by SDS-PAGE (CBB staining) (Fig.1). Endoand exo-DNase activities and RNase activity were not detected with 100 U/ml RNaseH in 50 ul reaction at 37℃.
Reagents Supplied with Enzyme
RNaseH Reaction Buffer (10 X): 100 mM Tris-HCl (pH 8.0), 100 mM MgCl2, 500 mM NaCl, 10 mM DTT, 500 ug/ml BSA (Bovine Serum Albumin)
*Caution
To avoide contamination of trace amounts of nucleic acids in BSA, use reaction buffer that does not contain BSA and use RNaseH at higher concentrations.
Data Link
UniProtKB/Swiss-Prot P0A7Y4 (RNH_ECOLI)
References
- Gubler U (1987) “Second-strand cDNA synthesis: mRNA fragments as primers.” Method Enzymol 152: 330-335 PMID: 3309563
- Sambrook J & Russell DW (2001) Molecular Cloning, Chapter 11 “Preparation of cDNA Libraries and Gene Identification”. CSHL Press
- Vournakis JN et al (1975) “Electrophoretic patterns of deadenylylated chorion and globin mRNAs.” Proc.Natl.Acad.Sci.USA 72: 2959-2963 PMID: 1059086
- Donis-Keller H (1979) “Site specific enzymatic cleavage of RNA.” Nucleic Acids Res. 7: 179-192 PMID: 386279
