Introduction
Bovine pancreatic deoxyribonuclease (also known as DNase) is an endonuclease. It acts on the phospholipid bond, especially the bond adjacent to pyrimidine nucleoside, to produce polynucleotides with free hydroxyl at the 3 'end and phosphate at the 5' end. The optimum pH value of is 7.8. DNase can be activated by divalent metals and inhibited by chelates such as EDTA and sodium dodecyl sulfate. 5 mM calcium ion can be used as a stabilizer to protect DNase from being decomposed by hydrolase. This product is extracted from bovine pancreas and prepared by chromatography to remove the pollution of other hydrolases.
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Details
Specifications
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Features
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Specifications
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Molecular weight
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32KDa
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Isoelectric point
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8.9
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pH range
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4.0 - 10.0
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Specific activity
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3000 kunitz units/mg
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Preservation conditions
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It is recommended to store at -20°C to ensure the stability of activity to the greatest extent (normal temperature transportation or storage will not reduce enzyme activity).
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Solution
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50mM Tris (pH7.4)，10mM CaCl2，50% (v/v) glycerol
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Recommended application
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Remove DNA
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RNase Residue
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Not detection
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RNase-Free DNase I Prepare
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Dissolve DNase I at 1 mg/ml in 0.1M iodoacetic acid plus 0.15M sodium acetate at a final pH of 5.3. The solution is then heated 40-60 minutes at 55°C and cooled. Finally, 1M CaCl2 is added to the solution to 5mM. Store frozen in small aliquots.
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Technical Validation
The DNase I Set was validated for RNase-free performance using bacterial RNA samples. After DNase I treatment, agarose gel electrophoresis showed intact RNA bands without visible RNA degradation, indicating no detectable RNase carryover under the tested conditions.
DNase activity was evaluated using liver tissue DNA extraction samples. Compared with the no-DNase and DNase Buffer control groups, samples treated with DNase I showed a strong reduction in DNA yield by NanoDrop measurement and no obvious genomic DNA band by agarose gel electrophoresis, supporting effective DNA digestion activity.
The effect of DNase I digestion on downstream qPCR detection was further tested using porcine blood samples spiked with HBV DNA. Control samples without DNase treatment showed Ct values around 30, while DNase-treated samples showed no Ct signal, indicating that DNA templates were effectively degraded and no detectable amplifiable DNA remained under the tested conditions.
Stability testing showed that liquid DNase I stored at room temperature for two weeks performed similarly to DNase I stored at 4°C in bacterial RNA extraction and tissue DNA digestion tests. These results support the use of the DNase I Set for on-column or magnetic bead-based RNA extraction workflows requiring reliable genomic DNA removal.
Ordering information
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Contents
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C12131
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C12132
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Dnase I, Lyophilizate, >3000 units/mg protein
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100 mg/Bottle
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1 g/Bottle
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