碱性磷酸酶

保存条件 ：Store at -5 to -30°C.

英文名 ：Anza™ Alkaline Phosphatase

库存 ：现货

供应商 ：恒斐生物

CAS号 ：相关应用： 限制性内切酶和克隆酶

规格 ：2000 reactions

Anza™ Alkaline Phosphatase描述

Invitrogen™ Anza™ Alkaline Phosphatase is intended for use in the removal of 3' and 5' phosphate groups that remain after Anza™ restriction enzyme digestion, such as for the dephosphorylation of vectors prior to insert ligation and removal of 5′ phosphates prior to end-labeling. Since the 5′ phosphate group is required for DNA ligation, treatment of vectors with Anza Alkaline Phosphatase prevents self-ligation and re-circularization, resulting in decreased vector background when cloning.

Features of Anza Alkaline Phosphatase:

• Validated for use with all Anza restriction enzymes
• Dephosphorylates in 15 minutes at 37°C
• Heat inactivated after 5 minutes at 80°C
• Fully active in Anza Buffer
• Dephosphorylates all types of DNA ends (blunt, 5'- and 3'-overhangs)

Flexible protocol
Some restriction enzymes prohibit or reduce the dephosphorylation process when digestion and dephosphorylation are performed simultaneously. Therefore, Anza Alkaline Phosphatase may be used in one of three ways depending upon the restriction enzyme used:

• One-Step Protocol: Simultaneous restriction enzyme digestion and dephosphorylation in Anza buffer
• Heat Inactivation Two-Step Protocol: Digestion is carried out first, followed by heat inactivation (20 minutes at 80oC) then dephosphorylation
• Column Purification Two-Step Protocol: Digestion is carried out first, followed by column purification then dephosphorylation

No need for enzyme removal
Anza Alkaline Phosphatase is completely inactivated after 5 minutes at 80°C, so enzyme removal via phenol/chloroform extraction or column purification is not required prior to subsequent ligation steps. If the One-Step protocol was used, heat the reaction mixture for 20 minutes at 80oC to inactivate the restriction enzyme as well as the alkaline phosphatase. Heat inactivation of the alkaline phosphatase prevents residual phosphatase from dephosphorylating the DNA fragments added to the reaction mixture for ligation.

Anza™ Alkaline Phosphatase is part of the Anza™ Restriction Enzyme Cloning System, unifying the traditional cloning processes. Other Anza DNA Modifying Enzymes include theAnza T4 DNA Ligase Master Mix, Anza T4 PNK Kit, Anza DNA Blunt End Kit, and Anza DNA End Repair Kit.