Details

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 for15 minutes

Assay Conditions

37 mM Tri-HCl (pH 8.0)14.8 mM KCl7.4 mM MgCl₂2.2 mM β-mercaptoethanol0.6 nmoles [32P]poly(A)·poly(dT)Reaction volume of 25 μl (6)

Storage Buffer

20 mM Tris-HCl (pH 7.5) 300 mM KCl1.0 mM DTT7 mM EDTA20 mM magnesium acetate 50% (v/v) glycerol

Storage Conditions

Store at -20°CProduct shipped on Dry Ice

Downloads

Certificate of Analysis -Current LotMSDS PDF-Current Lot

References

(1) Donis-Keller, H. (1979) Nucleic Acids Res. 7, 179-182(2) Okayama, H. and Berg, P. (1982) Mol. Cell Bio. 2, 161-170(3) Gubler, U. and Hoffman, B.J. (1983) Gene 25, 263-269(4) Keller, W. and Crouch, R. (1972) Proc. Natl. Acad. Sci. USA 69, 3360-3364(5) Goodwin, E.C. and Rottman, F.M. (1982) Nucl. Acids Res. 20, 916(6) Hillenbrand, G. and Staudenbauer, W.L. (1982) Nucl. Acids Res. 10, 833-852