RNase H in Trehalose

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 at 37°C.

Heat Inactivation

65ºC for 15 minutes

Assay Conditions

20 mM HEPES-KOH (pH 8.0)
50 mM KCl
10 mM MgCl₂
1 mM DTT
0.6 nmoles [³²P]poly(A)·poly(dT)
Reaction volume of 25 μl (6)

Storage Buffer

50 mM Tris-HCl (pH 7.5) 
300 mM KCl
1 mM DTT
7 mM EDTA
20 mM Magnesium Acetate
1 M Trehalose

Storage Conditions

Store at -20°C
Product shipped on Dry Ice