Description: NZY RNase H is a recombinant endoribonuclease purified from an Escherichia coli strain that carries the cloned RNase H gene (rnh). The enzyme specifically hydrolyses the phosphodiester bonds of RNA which is hybridized to DNA to produce- 5´ phosphate-terminated oligoribonucleotides and single-stranded DNA. RNase H does not degrade ss- and dsDNA or unhybridized RNA. It is a key enzyme in the removal of mRNA after first-strand cDNA synthesis. Treating cDNA with RNase H prior to PCR can improve sensitivity as bond RNA in the cDNA reaction may prevent binding of the amplification primers. RNase H treatment is often necessary when amplifying longer, full length cDNA targets.
Features:
– Specifically hydrolyses the phosphodiester bonds of RNA which is hybridized to DNA
– Can be inactivated in 10 min at 65 ºC
Applications:
– Removal of mRNA after first-strand cDNA synthesis
– Removal of poly(A) tails on mRNAs after hybridization with oligo(dT)
– Site-specific cleavage of RNA
Specifications:
| Optimal reaction temperature: | 37 ºC |
| Denaturing conditions: | The activity of NZY RNase H is inhibited by metal chelators (e.g. EDTA) and sulfhydryl SH-blocking reagents. Temperatures ≥ 65 °C inactivate the enzyme. |
| Storage conditions: | Store at -20 ºC |
| Shipping conditions: | Shipped on dry ice |
Components:
– NZY RNase H (E. coli) (5 U/μL)
– Reaction buffer (10x)