Note:This product needs to be packed with ice bags and shipped through FedEx or UPS.
It will arrive within 7–10 days. The shipping fee is included in the product price.
Introduction
Bovine Pancreatic Deoxyribonuclease (DNase I)
- Nature and Function:
- DNase I is an endonuclease enzyme extracted from bovine pancreas.
- It primarily acts on DNA, cleaving the phosphodiester bonds, especially those adjacent to a pyrimidine nucleoside.
- The cleavage results in oligonucleotides that terminate with a phosphate group at the 5′ end and a hydroxyl group at the 3′ end.
- Optimal Conditions and Stabilization:
- The optimum pH for DNase I activity is 7.8.
- It requires divalent metal ions for activation; common activators include magnesium and calcium ions.
- 5 mM calcium ions are particularly used to stabilize DNase I, protecting it from degradation by proteases.
- Inhibition:
- DNase I activity can be inhibited by chelating agents such as EDTA, which bind and remove divalent cations essential for its activity.
- Sodium dodecyl sulfate (SDS) also inhibits its activity, likely through interference with the enzyme’s interaction with DNA.
- Purification:
- The enzyme is purified through chromatographic techniques to remove contaminants, particularly other hydrolases.
RNase Free DNase I Set (For RNA Purification)
- Purpose:
- This kit is designed for the removal of DNA from RNA preparations in column or magnetic bead-based RNA extraction protocols.
- Procedure:
- Biological samples are initially lysed, and ethanol is added to optimize conditions for RNA binding to the column or magnetic beads.
- After binding, the sample undergoes a wash step to remove impurities.
- DNA Digestion:
- DNase I, along with a specific buffer, is applied to the RNA-bound membrane or beads.
- The mixture is incubated at room temperature (25-37 °C) for about 15 minutes, allowing DNase I to digest and remove any DNA present.
- Post-Digestion Processing:
- Following digestion, the column, or beads are washed to remove DNase and the degraded DNA fragments.
- Finally, RNA is eluted using DEPC-treated water to prevent any RNAse contamination.
Details
Specifications
Features | Specifications |
Molecular weight | 32KDa |
Isoelectric point | 8.9 |
pH range | 4.0–10.0 |
Specific activity | 10 kunitz units/μl |
Advantages
- Pure natural protein, from bovine pancreas
- High activity and purity, >10 kunitz units/μl
- High cost performance
- Completely remove RNase contamination
- Ready to use liquid type
- Ultra-high dosage (10μl/time,100μ/time) to remove more genomic DNA
Ordering information
Contents | C12133 | C12134 |
RNase-Free DNase I (10 units/μl) Plus DNase Buffer | 500 Preps (Plus 60 ml DNase Buffer) | 5000 Preps (Plus 600 ml DNase Buffer) |
Storage and Stability
Magen RNase-Free DNase I is stable for up to 1 year after delivery when stored at -20 °C.
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