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RNase H

Ribonuclease H from E.coli

Ribonuclease H is an endonuclease which specifically degrades the RNA moiety of a DNA:RNA hybrid, without affecting the unhybridized RNA. It does not affect double-stranded RNA or DNA. It is provided in a 50% glycerol storage buffer. It is also available in a Trehalose Buffer (Suitable for lyophilization).


RNase H IN GLYCEROL

Catalog No. Item Name Size/Volume Concentration Price (USD)
RNHG-50 RNase H in Glycerol 50 U 1000 U/mL $25.00
Quantity :
RNHG-250 RNase H in Glycerol 250 U 1000 U/mL $55.00
Quantity :
RNHG-1000 RNase H in Glycerol 1,000 U 1000 U/mL $150.00
Quantity :
RNHG-C RNase H in Glycerol Custom Custom Please inquire  


RNase H IN TREHALOSE

Catalog No. Description Size/Volume Concentration Price  
RNHT-50 RNase H in Trehalose 50 U 1000 U/mL $34.00
Quantity :
RNHT-250 RNase H in Trehalose 250 U 1000 U/mL $62.50
Quantity :
RNHT-1000 RNase H in Trehalose 1000 U 1000 U/mL $180.00
Quantity :
RNHT-C RNase H in Trehalose Custom Custom Please inquire  

Applications:

  • Enabling the synthesis of second-strand cDNA by removal of the RNA
  • Used in conjunction with AMV RT and T7 RNA Polymerase in amplification of RNA

Unit Definition:

One unit of RNaseH is that amount of enzyme which catalysis the production of one nmol of acid-soluble nucleotide in 20 minutes at 37ºC using the following reaction conditions:

  • 40 mM TrisHCl, pH 7.5
  • 1.0 µM [3H]-poly (rA):24 µM poly(dT)
  • 4.0 mM MgCl2
  • 1.0 mM DTT
  • 30 µg/mL BSA
  • 4.0% glycerol

Storage Buffer Conditions of RNaseH in Glycerol:

  • 20:0 mM TrisHCl, pH 7.5
  • 300 mM KCl
  • 20.0 mM Magnesium Acetate
  • 7.0 mM EDTA
  • 1.0 mM Dithiothreitol
  • 50% Glycerol
  • 0.2% Triton X100
  • 200.0 µg/ml BSA

Storage Buffer Conditions of RNaseH in Trehalose. Same as above, except for 1.0 M Trehalose instead of Glycerol.

Quality Control:

DNase Activity:

One-half µg of Hae fragments of Phi X-174 DNA is incubated at 37ºC with 2.5 units of RNase H for 3 hours, and then electrophoresed in a native agarose gel simultaneously with control positive DNase 1 reactions. No more than the equivalent of 2.5 X 10 E-4 unit of DNase 1 is detected.

Ribonuclease Activity:

One microgram of an RNA Ladder is incubated for 2 hours at 37ºC with 4.0 units of RNase H, and then electrophoresed in a native agarose gel simultaneously with control positive RNase 1A reactions. No more than the equivalent of 8 X 10-8 unit of RNase 1A is detected.

Specific Activity:

The specific activity of the E. coli RNase H is no less than 300,000 units per mg.

References:

  1. Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989) Molecular Cloning: A Laboratory Manual, (2nd Ed.), 8.64 – 8.65