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Introducción
Bovine Pancreatic Deoxyribonuclease (ADNasa 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′ fin.
- 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.
- Dodecil sulfato de sodio (Ficha de datos de seguridad (MSDS)) 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)
- Objetivo:
- This kit is designed for the removal of DNA from RNA preparations in column or magnetic bead-based extracción de ARN protocols.
- Procedimiento:
- 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:
- ADNasa 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 minutos, 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.
- Finalmente, RNA is eluted using DEPC-treated water to prevent any RNAse contamination.
Detalles
Especificaciones
Características | Especificaciones |
Peso molecular | 32kda |
Isoelectric point | 8.9 |
pH range | 4.0–10.0 |
Actividad específica | 10 kunitz units/μl |
Ventajas
- 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
Información sobre pedidos
Contenido | C12133 | C12134 |
RNase-Free DNase I (10 units/μl)
Plus DNase Buffer |
500 preparativos (Plus 60 ml DNase Buffer) | 5000 preparativos (Plus 600 ml DNase Buffer) |
Almacenamiento y estabilidad
Magen RNase-Free DNase I is stable for up to 1 year after delivery when stored at -20 °C.
Reseñas
Aún no hay reseñas.