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介紹
核糖核酸酶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′-相鄰嘧啶核苷酸的核糖. The resulting 2′,3′-cyclic phosphate is hydrolyzed to the corresponding 3′-核苷磷酸. The highest activity is exhibited with single-stranded RNA. RNase A is a single chain polypeptide containing 4 二硫橋.
A major application for RNase A is the removal of RNA from preparations of 質粒DNA. The enzyme is active under a wide range of reaction conditions. 低鹽濃度下 (0 到 100 毫摩爾氯化鈉), RNase A cleaves single-stranded and double-stranded RNA as well as the RNA strand in RNA-DNA hybrids. 然而, at NaCl concentrations of 0.3 M or higher, RNase A specifically cleaves single-stranded RNA.
細節
規格
特徵 | 規格 |
純度 | ≥60% RNase A basis (安全資料表-頁) |
Enzymatic activity | >50 庫尼茨單位/毫克蛋白質 |
Optimum reaction temperature | 60℃ (effective active temperature is 15-70°C) |
運輸條件 | Normal temperature transportation |
保存條件 | -20-8℃, 幹儲存, 長期存儲應放置在-20°C下. |
應用 1: adding in the extraction process | 1. Plasmid Extraction: add RNase A (25毫克/毫升) to buffer P1 with a final concentration of 100-300μg/ml.
2. DNA萃取: add RNase A (25毫克/毫升) to the digestion solution with a final concentration is 100-400μg/ml, mix well and place at room temperature for 10-15 分分鐘. when SDS/CTAB in lysate exceeds 2%, RNase activity will be significantly inhibited; 胍 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. |
應用 2: | 1. Remove RNA contamination from crude genomic DNA products: add DNase free RNase A (10毫克/毫升) to crude DNA products with a final concentration of 10-100μg/ml. 混合後, 放在室溫下 10 分分鐘.
2. Remove RNA contamination from plasmid DNA products: add DNase free RNase A (10毫克/毫升) 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 分分鐘. |
訂購資訊
內容 | C12123 | C12124 | C12128 | C12129 |
RNase A Solution (25毫克/毫升) | 10 毫升 | 100 毫升 | ||
DNase Free RNase A Solution (10毫克/毫升) | 10 毫升 | 100 毫升 |
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