Set dNase

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Uvod

Bovine Pancreatic Deoxyribonuclease (DNaza I)

1. 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.
2. 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.
3. Inhibition:
   • DNase I activity can be inhibited by chelating agents such as EDTA, which bind and remove divalent cations essential for its activity.
   • Natrijev dodecil sulfat (SDS) also inhibits its activity, likely through interference with the enzyme’s interaction with DNA.
4. Purification:
   • The enzyme is purified through chromatographic techniques to remove contaminants, particularly other hydrolases.

RNase Free DNase I Set (For RNA Purification)

1. Svrha:
   • This kit is designed for the removal of DNA from RNA preparations in column or magnetic bead-based RNA extraction protocols.
2. Postupak:
   • 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.
3. DNA Digestion:
   • DNaza 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 minuta, allowing DNase I to digest and remove any DNA present.
4. 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.

pojedinosti

Tehnički podaci

Prednosti

• Pure natural protein, from bovine pancreas
• High activity and purity, >10 kunitz units/μl
• Visoko troškovni učinak
• Completely remove RNase contamination
• Ready to use liquid type
• Ultra-high dosage (10μl/time,100μ/time) to remove more genomic DNA

Informacije o naručivanju

Skladištenje i stabilnost

Magen RNase-Free DNase I is stable for up to 1 year after delivery when stored at -20 °C.