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Einführung
Characteristics of RNase A:
- Enzyme Type:
- RNase A is an endoribonuclease that specifically degrades single-stranded RNA.
- Specificity:
- It specifically cleaves RNA at cytosine (C) and uracil (U) residues.
- Action Mechanism:
- The enzyme cleaves the phosphodiester bonds between the 5′-ribose of a nucleotide and the phosphate group attached to the 3′-ribose of an adjacent pyrimidine nucleotide.
- This action initially forms a 2′,3′-cyclic phosphate, which is subsequently hydrolyzed to a 3′-nucleoside phosphate.
- Structure:
- RNase A is a single-chain polypeptide that includes 4 disulfide bridges, contributing to its stability.
- Optimal Activity:
- The highest enzymatic activity of RNase A is observed when acting on single-stranded RNA.
Applications of RNase A:
Primary Use:
- RNase A is extensively used to remove RNA from preparations of Plasmid-DNA, ensuring that the DNA samples are free of RNA contamination.
Activity Across Different Conditions:
Salt Concentration Influence:
- At low salt concentrations (0 Zu 100 mM NaCl), RNase A can cleave both single-stranded and double-stranded RNA, and the RNA strand in RNA-DNA hybrids.
- At higher salt concentrations (0.3 M NaCl or more), its activity becomes specific to single-stranded RNA.
Einzelheiten
Spezifikationen
Merkmale | Spezifikationen |
Aussehen | White or light yellow lyophilized powder |
Molecular weight | 13.7 KDa |
Reinheit | ≥60% RNase A (Sicherheitsdatenblatt-SEITE) |
Specific activity | >50 Kunitz units/mg protein |
Aufbewahrungsbedingungen | -20-8 °C. RNase is stable under heating and detergent conditions and can withstand high temperature treatment of 100 °C. |
DNase residue detection | Nicht erkannt. In 10μl (einschließlich 200 ng plasmid) Lösung, RNase A was added to the final concentration of 50-100 μg/ml and placed at room temperature for 30 Protokoll. Nach der Elektrophorese, the main band was obvious without degradation. |
Anwendung | Remove RNA from preparations of plasmid DNA |
Vorteile
- High activity, >50 U/mg
- High cost performance
- Hohe Reinheit, low impurity protein pollution
- No residual Dnase or trace Dnase
Bestellinformationen
Inhalt | C12120 | C12121 |
RNase A, Lyophilisat, >50 U/mg of protein | 1 G | 10 G |
Lagerung und Stabilität
Magen recommends storage at -20-8 °C. When stored at 2-8 °C, the product retains activity for at least 2 Jahre.
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