M.B. Enzymes GmbH

DNase I, RNase free.

Description

Deoxyribonuclease I (EC 3.1.21.1) is isolated from bovine pancreas catalyzes the hydrolysis of dsDNA to produce 3´-hydroxyl oligonucleotides; at high concentration of the enzyme, ssDNA will also be digested. In the presence of Mg²⁺ DNase I produces nicks in dsDNA, while in the presence of Mn²⁺ the enzyme produces ds breaks. M.W. approx. 37000. Specific activity approx. 3000 U/mg protein.

Unit definition

One unit will cause an increase in absorbance of calf thymus DNA solution at a rate of 0,001 A₂₆₀ units/min/ml at 25°C and pH 5.0.

Reaction conditions

50 mM Tris-HCl pH 7.5, 10 mM MnCl₂ (or 10 mM MgCl₂in nick translation protocols), 50 µg/ml BSA, 1 µg DNA, 0,1-2U of the enzyme (degradative use) or 20 mU(nick translation) in a 100µl reaction volume. Incubate at 37°C for 60 min. Stop reaction by adding EDTA to 25 mM (end conc.).

Storage

Store at -20°C.

Applications

nick translation; DNase footprinting, preparation of DNA-free RNA (RT-PCR, in vitro transcription/translation systems), cloning random DNA fragments by catalyzing ds cleavage of DNA in the presence of Mn²⁺.

Quality control

activity, SDS-PAGE purity, no contaminating ribonuclease activity.

References

1. Kunitz, M. (1950) J. gen. Physiol. 33, 363. 2. Moor, S. (1981) In The Enzymes, vol 14A, 281-298. 3. Campbel, V.W., Jackson, D.A. (1980) J. Biol. Chem. 255, 3726-3735.

Catalog #

M15.5

M15.7

Pack size

2.500 U

10.000 U

DNase I, RNase free.
Description	Deoxyribonuclease I (EC 3.1.21.1) is isolated from bovine pancreas catalyzes the hydrolysis of dsDNA to produce 3´-hydroxyl oligonucleotides; at high concentration of the enzyme, ssDNA will also be digested. In the presence of Mg²⁺ DNase I produces nicks in dsDNA, while in the presence of Mn²⁺ the enzyme produces ds breaks. M.W. approx. 37000. Specific activity approx. 3000 U/mg protein.
Unit definition	One unit will cause an increase in absorbance of calf thymus DNA solution at a rate of 0,001 A₂₆₀ units/min/ml at 25°C and pH 5.0.
Reaction conditions	50 mM Tris-HCl pH 7.5, 10 mM MnCl₂ (or 10 mM MgCl₂in nick translation protocols), 50 µg/ml BSA, 1 µg DNA, 0,1-2U of the enzyme (degradative use) or 20 mU(nick translation) in a 100µl reaction volume. Incubate at 37°C for 60 min. Stop reaction by adding EDTA to 25 mM (end conc.).
Storage	Store at -20°C.
Applications	nick translation; DNase footprinting, preparation of DNA-free RNA (RT-PCR, in vitro transcription/translation systems), cloning random DNA fragments by catalyzing ds cleavage of DNA in the presence of Mn²⁺.
Quality control	activity, SDS-PAGE purity, no contaminating ribonuclease activity.
References	1. Kunitz, M. (1950) J. gen. Physiol. 33, 363. 2. Moor, S. (1981) In The Enzymes, vol 14A, 281-298. 3. Campbel, V.W., Jackson, D.A. (1980) J. Biol. Chem. 255, 3726-3735.
Catalog #	M15.5	M15.7
Pack size	2.500 U	10.000 U