Thermo Scientific™ RNase H (5 U/μL)

• Does not hydrolyze the phosphodiester bonds within single-stranded and double-stranded DNA and RNA

• The absence of endo-, exodeoxyribonucleases and ribonucleases confirmed by appropriate quality tests

Source:

• E.coli MRE-600 cells

Molecular Weight:

• 18.4kDa monomer

Definition of Activity Unit:

• One unit of the enzyme catalyzes the formation of 1nmol of acid soluble products in 20 min. at 37°C
• Enzyme activity is assayed in the following mixture: 20mM Tris-HCl (pH 7.8), 40mM KCl, 8mM MgCl₂, 1mM DTT, 24μM [³H]-poly(A)·poly(dT), 0.03mg/mL BSA, 4% (v/v) glycerol

• The enzyme is supplied in: 25mM HEPES-KOH (pH 8.0), 50mM KCl, 0.1mM EDTA, 1mM DTT, 0.1mg/mL BSA and 50% (v/v) glycerol

10X Reaction Buffer:

• 200mM Tris-HCl (pH 7.8), 400mM KCl, 80mM MgCl₂, 10mM DTT

Inhibition and Inactivation:

• Inhibitors: metal chelators, SH-blocking reagents
• Inactivated by heating at 65°C for 10 min.

Recommended for:

• Removal of mRNA prior to synthesis of second strand cDNA (1)
• RT-PCR and qRT-PCR: removal of RNA after first strand cDNA synthesis
• Removal of the poly(A) sequences of mRNA after hybridization with oligo(dT) (2)
• Site-specific cleavage of RNA (3)
• Studies of in vitro polyadenylation reaction products