计算溶液所需的质量、体积或浓度。
虾碱性磷酸酶 (Shrimp Alkaline Phosphatase,rSAP) 催化 DNA 和 RNA 的 5’和 3’磷酸单脂的去磷酸化反应。该磷酸酶在分子生物学研究中应用广泛,如去除 DNA 和 RNA 末端的磷酸基团,用于后续的克隆和探针末端标记。在克隆中,可使线性载体去磷酸化,防止自连。虾碱性磷酸酶在65℃ 温育 5 分钟即可完全的、不可逆的失活,因此,在连接或末端标记之后,可以不用去除热敏磷酸酶。基于以上特性,该酶是传统去磷酸化酶--小牛肠碱性磷酸酶(CIP)的绝佳替代物。
组分表:
| 组分货号 | 组分名称 | 物理外观 | 500U | 5×500U | 储存 |
| S1438762A | 虾碱性磷酸酶 (2U/μl) | 液体 | 250μl | 5×250μl | -20℃,避免反复冻融。 |
| S1438762B | 10×SAP 缓冲液 | 液体 | 1ml | 5×1ml | -20℃,避免反复冻融。 |
应用:
1. DNA 和 RNA 的去磷酸化;
2. 防止克隆载体的自连;
3. 制备 5´末端标记模板。
使用方法(核酸末端去磷酸化):
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注意:1 pmol 的 5' 末端 DNA,约等于 1 μg 单酶切后的 3 kb 质粒。
Shrimp Alkaline Phosphatase (rSAP) catalyzes the dephosphorylation of 5′ and 3′ phosphate monoesters on DNA and RNA. It is widely used in molecular biology research, such as removing phosphate groups from the termini of DNA and RNA for subsequent cloning and probe end-labeling. For cloning applications, it dephosphorylates linear vectors to prevent self-ligation. rSAP can be fully and irreversibly inactivated by incubation at 65 °C for 5 minutes, so purification of the thermolabile phosphatase is not required prior to ligation or end-labeling. Owing to these properties, this enzyme serves as an excellent alternative to the conventional dephosphorylation enzyme, Calf Intestinal Alkaline Phosphatase (CIP).
Components List:
| Component No. | Component Name | Appearance | 500U | 5×500U | Storage |
| S1438762A | Shrimp Alkaline Phosphatase (2U/μl) | Liquid | 250μl | 5×250μl | -20℃. Avoid freeze/thaw cycle. |
| S1438762B | 10×SAP Buffer | Liquid | 1ml | 5×1ml | -20℃. Avoid freeze/thaw cycle. |
Applications:
1. Dephosphorylation of DNA and RNA;
2. Prevention of self-ligation of cloning vectors;
3. Preparation of templates for 5′ end-labeling.
Protocol (Nucleic acid terminal dephosphorylation):
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Note: 1 pmol of 5′-DNA ends is approximately equivalent to 1 μg of linearized 3 kb plasmid after single restriction digestion.
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