RNase H

Source

E. coli

Applications

• Hybridization of a synthetic DNA oligomer to a complementary single-stranded region of a RNA molecule can be used to create a site that can be cleaved by RNase H (1)
• Used to remove RNA strand before second strand cDNA synthesis (2, 3)
• Ribonuclease H (RNase H) detects RNA-DNA regions in naturally occurring double-stranded DNA (4)
• Used to analyze in vitro polyadenylation reaction products (5)

Reagents Supplied

10X RNase H Reaction Buffer

Unit Definition

One unit is the amount of enzyme required to produce 1 nmol of acid soluble ribonucleotide from [32P]poly(A)·poly(dT) at 37°C in 20 minutes

Heat Inactivation

65ºC for 15 minutes

Assay Conditions

37 mM Tri-HCl (pH 8.0)
14.8 mM KCl
7.4 mM MgCl₂
2.2 mM β-mercaptoethanol
0.6 nmoles [32P]poly(A)·poly(dT)
Reaction volume of 25 μl (6)

Storage Buffer

20 mM Tris-HCl (pH 7.5) 
300 mM KCl
1.0 mM DTT
7 mM EDTA
20 mM magnesium acetate
50% (v/v) glycerol

Storage Conditions

Store at -20°C
Product shipped on Dry Ice