DNase I, or Deoxyribonuclease I, is an endonuclease that can digest single-stranded or double-stranded DNA. It hydrolyzes phosphodiester bonds to produce monodeoxynucleotides and oligodeoxynucleotides containing 5'-phosphate groups and 3'-OH groups. The optimal working pH range of DNase I is 7-8. The activity of DNase I depends on Ca2+ and can be activated by divalent metal ions such as Co2+, Mn2+, and Zn2+. In the presence of Mg2+, DNase I can randomly cut any site of double-stranded DNA; in the presence of Mn2+, DNase I can cut the DNA double strand at the same site, forming a blunt end or a sticky end with 1-2 nucleotides protruding, which can be used for the treatment of various RNA samples. Features Recombinant DNAse I from yeast RNase-free High enzymatic cleavage efficiency. More suitable for applications sensitive to RNase. Applications Removal of contaminating genomic DNA from RNA samples Degradation of DNA templates in transcription reactions Removal of gDNA before RNA extraction or reverse transcription. Removal of template DNA in in vitro transcription. Specifications Cat.No. 14549ES80 / 14549ES90 Size 1000 U / 5000 U Unit definition: Using calf thymus DNA as substrate, at 25℃, pH 5.0, the amount of enzyme required to increase the absorbance of the reaction solution at 260 nm by 0.001 within 1 min is defined as 1 activity unit (Kunitz Unit). Source Pichia yeast strain carrying the gene cloned from bovine pancreatic DNase I Purity ≥95% Reaction conditions 37℃, 15-30 min Heat inactivation conditions Add 2.5 mM EDTA solution and mix well, then incubate at 65℃ for 10 min Components Components No. Name 14549ES80 14549ES90 14549-A Recombinant DNase I (RNase-free, Yeast) (5 U/μL) 200 μL 1 mL 14549-B DNase I Reaction Buffer (10×) 1 mL 5×1 mL Figures 1. Strong digestive activity Figure 1. Plasmid DNA digestion 1 μg of plasmid DNA was digested using different amounts of DNase I from Yeasen and Supplier A. The results showed that the plasmid DNA removal efficiency of 14549ES was better than that of Supplier A. 2. No RNase residue Figure 2. RNase residue detection 10 U of Yeasen Recombinant DNase I (RNase-free, Yeast) was incubated with RNA substrate at 37°C for 1 h. Agarose gel electrophoresis analysis showed that no residual RNase activity was detected in the three batches of the enzyme preparation, completely eliminating the risk of RNA sample degradation. 3. RNA extraction application verification Figure 3. RNA extraction application verification 20 U of the enzyme preparation was used to treat 12 pre-treatment samples of mouse liver from different sources. RNA extraction and agarose gel electrophoresis analysis showed that the RNA integrity of the enzyme-treated group was good, indicating that this DNase I can effectively digest DNA without affecting the quality of RNA, and fully meets the technical requirements of RNA extraction experiments.
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