SKU: 19449462551

DNase Ⅰ

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Description

DNase ⅠProduct Specification Species Bovine Pancreatic Synonyms DNASE,Deoxyribonuclease 1 Expression System E. coli Molecular Weight 72kDa (Reducing) Purity >95% by SDS PAGE&HPLC Conjugation Unconjugated Tag His Tag, MBP Tag Physical Appearance Liquid Storage Buffer 10 mM Tris HCl, 2 mM CaCl2 ,50% Glycerol(pH 7. 6, 25C) Stability & Storage Store at 25 ~ 15 for 2 years Reference [1] Vanecko S, Laskowski M. Studies of the Specificity of Deoxyribonuclease I[J].

Product Specification


Species Bovine Pancreatic
Synonyms DNASE,Deoxyribonuclease-1
Expression System E.coli
Molecular Weight

72kDa (Reducing)

Purity >95% by SDS-PAGE&HPLC
Conjugation Unconjugated
Tag His Tag, MBP Tag
Physical Appearance Liquid
Storage Buffer

10 mM Tris-HCl, 2 mM CaCl2 ,50% Glycerol,(pH 7.6, 25°C)

Stability & Storage

Store at -25 ~ -15℃ for 2 years

Reference

[1] Vanecko S, Laskowski M. Studies of the Specificity of Deoxyribonuclease I[J]. Journal of Biological Chemistry, 1961, 236(236):3312-6.

[2] Kienzle N, Young D, Zehntner S, et al. DNaseI treatment is a prerequisite for the amplification of cDNA from episomal-based genes[J]. Biotechniques, 1996, 20(4):612-6.

[3] Michael,R, Green, et al. Human β-globin pre-mRNA synthesized in vitro is accurately spliced in xenopus oocyte nuclei[J].Cell, 1983, 32(3):681-694.

Background

DNase I (Deoxyribonuclease I), can digest single or double-stranded DNA to produce mono deoxynucleotides or single or double-stranded oligo deoxynucleotides, its optimal working pH range is 7-8. DNase I activity is dependent on Ca2+ and can be activated by other bivalent metal ions such as Mg2+, Mn2+, Zn2+, etc. In the presence of Mg2+, the enzyme can randomly recognize and cut any site on any strand of DNA. In the presence of Mn2+, two strands of DNA can be cut at the same site to form sticky ends with flat ends or 1-2 nucleotides protruding.

Components

Storage Solution:  2 U/ul DnaseⅠ、10mM Tris-Hcl、2mM  CaCl2、50%Glycerol (pH7.6, 25℃)

10*Reaction Buffer: 100mM Tris-Hcl、25mM  MgCl2、5mM  CaCl2 (pH7.6, 25℃)

Protocol

This step is suitable for linearization of 1 μg DNA (≥100 nt) and can be scaled up according to experimental needs.

1)Add the following components in sequence

Components

Volume

Plasmid DNA

1μg DNA

10*Reaction Buffer

2μl

DnaseⅠ (2U/μl)

1μl

RNase-free ddH2O

Up to 50μl

2)Incubate at 37°C 1 h.

Guidelines

1. EDTA should be added to a final concentration of 5 mM to protect RNA from being degraded during enzyme inactivation 2. Please avoid repeated freeze-thaw cycles

Unit Definition

One unit is defined as the amount of enzyme which will completely degrade 1 µg of pBR322 DNA in 10 minutes at 37°C in DNase I Reaction Buffer.
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SKU: 19449462551

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