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Einführung
RNase A is an endoribonuclease that specifically degrades single-stranded RNA at C and U residues. It cleaves the phosphodiester bond between the 5′-ribose of a nucleotide and the phosphategroup attached to the 3′-ribose of an adjacent pyrimidine nucleotide. The resulting 2′,3′-cyclic phosphate is hydrolyzed to the corresponding 3′-nucleoside phosphate. The highest activity is exhibited with single-stranded RNA. RNase A is a single chain polypeptide containing 4 disulfide bridges.
A major application for RNase A is the removal of RNA from preparations of Plasmid-DNA. The enzyme is active under a wide range of reaction conditions. At low salt concentrations (0 Zu 100 MM NaCl), RNase A cleaves single-stranded and double-stranded RNA as well as the RNA strand in RNA-DNA hybrids. Jedoch, at NaCl concentrations of 0.3 M or higher, RNase A specifically cleaves single-stranded RNA.
Einzelheiten
Spezifikationen
| Merkmale | Spezifikationen |
| Reinheit | ≥60% RNase A basis (Sicherheitsdatenblatt-SEITE) |
| Enzymatic activity | >50 Kunitz units/mg protein |
| Optimum reaction temperature | 60°C (effective active temperature is 15-70°C) |
| Transportbedingungen | Normal temperature transportation |
| Aufbewahrungsbedingungen | -20-8°C, trocken lagern, Die Langzeitlagerung sollte bei -20 °C erfolgen. |
| Anwendung 1: adding in the extraction process | 1. Plasmid Extraction: add RNase A (25mg/ml) to buffer P1 with a final concentration of 100-300μg/ml.
2. DNA-Extraktion: add RNase A (25mg/ml) to the digestion solution with a final concentration is 100-400μg/ml, mix well and place at room temperature for 10-15 Protokoll. when SDS/CTAB in lysate exceeds 2%, RNase activity will be significantly inhibited; Guanidin Salz(>4M guanidine hydrochloride or >3M guanidine isothiocyanate) also significantly inhibited RNase A. When RNase A is added to the lysate, the RNase digestion effect can be extracted by appropriately diluting to reduce the concentration of SDS, CTAB and guanidine salt. |
| Anwendung 2: | 1. Remove RNA contamination from crude genomic DNA products: add DNase free RNase A (10mg/ml) to crude DNA products with a final concentration of 10-100μg/ml. Nach dem Mischen, Bei Raumtemperatur legen für 10 Protokoll.
2. Remove RNA contamination from plasmid DNA products: add DNase free RNase A (10mg/ml) to crude DNA products with a final concentration of 10μg/ml. Nach dem Mischen, it can be directly used for sequencing at room temperature for 10 Protokoll. |
Bestellinformationen
| Inhalt | C12123 | C12124 | C12128 | C12129 |
| RNase A Solution (25mg/ml) | 10 ml | 100 ml | ||
| DNase Free RNase A Solution (10mg/ml) | 10 ml | 100 ml |
Rezensionen
Es gibt noch keine Rezensionen.