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導入
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′-隣接するピリミジンヌクレオチドのリボース. The resulting 2′,3′-cyclic phosphate is hydrolyzed to the corresponding 3′-ヌクレオシドリン酸. The highest activity is exhibited with single-stranded RNA. RNase A is a single chain polypeptide containing 4 ジスルフィドブリッジ.
A major application for RNase A is the removal of RNA from preparations of プラスミドDNA. The enzyme is active under a wide range of reaction conditions. 低塩濃度で (0 に 100 mm naCl), RNase A cleaves single-stranded and double-stranded RNA as well as the RNA strand in RNA-DNA hybrids. しかし, at NaCl concentrations of 0.3 M or higher, RNase A specifically cleaves single-stranded RNA.
詳細
仕様
| 特徴 | 仕様 |
| 純度 | ≥60% RNase A basis (SDS-ページ) |
| Enzymatic activity | >50 Kunitzユニット/Mgタンパク質 |
| Optimum reaction temperature | 60℃ (effective active temperature is 15-70°C) |
| 輸送条件 | Normal temperature transportation |
| 保存条件 | -20-8℃, 乾式保管, 長期保管は-20℃で保存してください. |
| 応用 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抽出: 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 分. when SDS/CTAB in lysate exceeds 2%, RNase activity will be significantly inhibited; Guanidine 塩(>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. |
| 応用 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. 混合後, 室温に置いてください 10 分.
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. 混合後, it can be directly used for sequencing at room temperature for 10 分. |
注文情報
| コンテンツ | C12123 | C12124 | C12128 | C12129 |
| RNase A Solution (25mg/ml) | 10 ミリリットル | 100 ミリリットル | ||
| DNase Free RNase A Solution (10mg/ml) | 10 ミリリットル | 100 ミリリットル |
レビュー
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