Invitrogen抗体|thermo46300018*

Invitrogen抗体|thermo46300018*

描述
T4 DNA连接酶在具有3'羟基和5'磷酸末端的双链DNA之间的ATP存在下催化磷酸二酯键的形成。*的T4 DNA连接酶缓冲液可优化连接，可在5分钟内完成。单链核酸不是该酶的底物。提供T4 DNA连接酶技术公告。

应用：克隆（钝端或粘性末端连接）。将接头或接头添加到平端DNA。

来源：从大肠杆菌中纯化 - 溶原菌NM989。

性能和质量测试：内切脱氧核糖核酸酶，3'和5'外切脱氧核糖核酸酶测定;结扎效率测试。

单位定义：一个单位催化在20分钟内将1nmol 32P标记的焦磷酸盐交换成ATP。在37°C。 （一个单位相当于大约300个粘性末端结扎单位。）

单位反应条件：66mM Tris-HCl（pH 7.6），6.6mM MgCl 2,10mM DTT，66μMATP，3.3μM32P标记的焦磷酸盐和0.1ml的酶，持续20分钟。在37°C。

Invitrogen抗体|thermo46300018*

Description

T4 DNA Ligase catalyzes the formation of phosphodiester bonds in the presence of ATP between double-stranded DNAs with 3´ hydroxyl and 5´ phosphate termini. The unique T4 DNA Ligase buffer optimizes ligation, which can be performed in 5 minutes (1). Single-stranded nucleic acids are not substrates for this enzyme. A T4 DNA Ligase Technical Bulletin is available.

Applications: Cloning (blunt-end or cohesive-end ligation) (2). Adding linkers or adapters to blunt-ended DNA (2).

Source: Purified from E. coli œ lysogen NM989.

Performance and Quality Testing: Endodeoxyribonuclease, 3´ and 5´ exodeoxyribonuclease assays; ligation efficiency tested.

Unit Definition: One unit catalyzes the exchange of 1 nmol 32P-labeled pyrophosphate into ATP in 20 min. at 37°C. (One unit is equal to approximately 300 cohesive-end ligation units.)

Unit Reaction Conditions: 66 mM Tris-HCl (pH 7.6), 6.6 mM MgCl2 , 10 mM DTT, 66 µM ATP, 3.3 µM 32 P-labeled pyrophosphate, and enzyme in 0.1 ml for 20 min. at 37°C.