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소개
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′ 끝.
- 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.
- 도데실황산나트륨 (SDS) also inhibits its activity, likely through interference with the enzyme’s interaction with DNA.
- 정화:
- The enzyme is purified through chromatographic techniques to remove contaminants, particularly other hydrolases.
RNase Free DNase I Set (For RNA Purification)
- 목적:
- This kit is designed for the removal of DNA from RNA preparations in column or magnetic bead-based RNA 추출 protocols.
- 절차:
- 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 ℃) for about 15 분, 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.
- 마지막으로, RNA is eluted using DEPC-treated water to prevent any RNAse contamination.
세부
명세서
특징 | 명세서 |
분자 무게 | 32KDa |
등전 지점 | 8.9 |
pH 범위 | 4.0–10.0 |
특정 활동 | 10 kunitz units/μl |
장점
- Pure natural protein, from bovine pancreas
- High activity and purity, >10 kunitz units/μl
- 높은 비용 성능
- Completely remove RNase contamination
- Ready to use liquid type
- Ultra-high dosage (10μl/time,100μ/time) to remove more genomic DNA
주문정보
내용물 | C12133 | C12134 |
RNase-Free DNase I (10 units/μl)
Plus DNase Buffer |
500 준비 (을 더한 60 ml DNase Buffer) | 5000 준비 (을 더한 600 ml DNase Buffer) |
보관 및 안정성
Magen RNase-Free DNase I is stable for up to 1 year after delivery when stored at -20 ℃.
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