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DNase I

Deoxyribonuclease I (DNase I) is DNA-specific endonuclease that cleaves both single-stranded DNA,double-stranded DNA and DNA-RNA hybrids, yielding 5'-phosphate-terminated polynucleotides with a free hydroxyl group on position 3' producing tetranucleotides. The activity of DNase I is strictly dependent on Ca2+ and can be activated by divalent metal ions such as Mg2+ or Mn2+. In the presence of Mg2+, DNase I nonspecifically recognizes and cleaves a double-stranded DNA at anysite on either strand, and in the presence of Mn2+, it recognizes and cleaves almost the same sites on both strands of the DNA to produce DNA fragments with blunt ends or sticky ends with 1~2 nucleotide overhangs. GenScript is offering DNase I produced by expression in a P. pastoris strain carrying a plasmid encoding the bovine DNase I.
$65.00
E00053-1000

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Description

Deoxyribonuclease I (DNase I) is DNA-specific endonuclease that cleaves both single-stranded DNA,double-stranded DNA and DNA-RNA hybrids, yielding 5'-phosphate-terminated polynucleotides with a free hydroxyl group on position 3' producing tetranucleotides. The activity of DNase I is strictly dependent on Ca2+ and can be activated by divalent metal ions such as Mg2+ or Mn2+. In the presence of Mg2+, DNase I nonspecifically recognizes and cleaves a double-stranded DNA at anysite on either strand, and in the presence of Mn2+, it recognizes and cleaves almost the same sites on both strands of the DNA to produce DNA fragments with blunt ends or sticky ends with 1~2 nucleotide overhangs.

GenScript is offering DNase I produced by expression in a P. pastoris strain carrying a plasmid encoding the bovine DNase I.

Note

For laboratory research use only. Direct human use, including taking orally and injection and clinical use are forbidden.

Properties
Source Recombinant DNase I expressed by yeast.
Species Bovine
Molecular Weight 35-40 kDa, on SDS-PAGE under reducing conditions.
Unit Definition One unit is the amount of the enzyme that increases the absorbance at 260 nm by 0.001 per minute at 25 °C, pH 5.0, with calf thymus DNA as the substrate.
Optimal active temperature 37 °C
Formulation Supplied as a solution of 20 mM sodium acetate, 5 mM CaCl2, 0.1 mM PMSF, 50% (v/v) glycerol, pH 6.5 at 25 °C.
Inactivation Add EDTA with a final concentration of 2.5 mM and heat the solution at 65 °C for 10 min can inactivate DNase I.
Storage & Stability This product remains stable up to 12 months at -20 °C. Avoid repeated freeze-thaw cycles.
Quality Control Specifications
AssaySpecifications
AppearanceClear, colorless liquid
Purity≥ 95% as analyzed by SDS-PAGE
Enzyme Activity≥ 2 U/μl
Endotoxin Level≤ 0.1 EU/μg of protein by gel clotting method
Residual RNaseNon-detectable
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Applications
·   DNA template digestion following in vitro transcription 
·   Genomic DNA digestion prior to RT-PCR
·   Preparation of DNA-free RNA samples
·   Nick-translation
·   Studies of DNA-protein interactions by DNase I, RNase-free footprinting·   Prevent cell clumping without affecting cell viability
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DNase I

Lane 1: Input Lane 7: 0.02 U
Lane 2: 1 U Lane 8: 0.01 U
Lane 3: 0.5 U Lane 9: 0.005 U
Lane 4: 0.2 U Lane 10: 0.002 U
Lane 5: 0.1 U Lane 11: 0.001 U
Lane 6: 0.05 U Lane 12: 0 U

Incubate with 100 ng plasmid DNA at 37 °C for 30 min, tiny amount of GenScript DNase I can completely digest the DNA. »

DNase I

Lane 1: IVT RNA (untreated)
Lane 2: IVT RNA + 2 U DNase I


Add 2 U of DNase I (E00053) to 20 μl in vitro transcription system, incubate at 37 °C for 30 min, GenScript DNase I can completely digest the DNA template. »

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Background
References 1.      Kienzle, N., et al. "DNaseI treatment is a prerequisite for the amplification of cDNA from episomal-based genes." BioTechniques 20.4 (1996): 612-616.
2.      Anderson, Stephen. "Shotgun DNA sequencing using cloned DNase I-generated fragments." Nucleic acids research 9.13 (1981): 3015-3027.
3.      Green, Michael R., Tom Maniatis, and D. A. Melton. "Human β-globin pre-mRNA synthesized in vitro is accurately spliced in Xenopus oocyte nuclei." Cell 32.3 (1983): 681-694.

--> 1.      Kienzle, N., et al. "DNaseI treatment is a prerequisite for the amplification of cDNA from episomal-based genes." BioTechniques 20.4 (1996): 612-616.
2.      Anderson, Stephen. "Shotgun DNA sequencing using cloned DNase I-generated fragments." Nucleic acids research 9.13 (1981): 3015-3027.
3.      Green, Michael R., Tom Maniatis, and D. A. Melton. "Human β-globin pre-mRNA synthesized in vitro is accurately spliced in Xenopus oocyte nuclei." Cell 32.3 (1983): 681-694.
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