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Introduction
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 to 100 mM NaCl), RNase A cleaves single-stranded and double-stranded RNA as well as the RNA strand in RNA-DNA hybrids. However, at NaCl concentrations of 0.3 M or higher, RNase A specifically cleaves single-stranded RNA.
Details
Specifications
| Features | Specifications |
| Purity | ≥60% RNase A basis (SDS-PAGE) |
| Enzymatic activity | >50 Kunitz units/mg protein |
| Optimum reaction temperature | 60°C (effective active temperature is 15-70°C) |
| Transportation conditions | Normal temperature transportation |
| Preservation conditions | -20-8°C, dry storage, long-term storage should be placed at -20°C. |
| Application 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 extraction: 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 minutes. when SDS/CTAB in lysate exceeds 2%, RNase activity will be significantly inhibited; Guanidine salt(>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. |
| Application 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. After mixing, place at room temperature for 10 minutes. 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. After mixing, it can be directly used for sequencing at room temperature for 10 minutes. |
Ordering information
| Contents | C12123 | C12124 | C12128 | C12129 |
| RNase A Solution (25mg/ml) | 10 ml | 100 ml | ||
| DNase Free RNase A Solution (10mg/ml) | 10 ml | 100 ml |
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