Invitrogen抗體|thermo46300018*

描述
T4 DNA連接酶在具有3'羥基和5'磷酸末端的雙鏈DNA之間的ATP存在下催化磷酸二酯鍵的形成。*的T4 DNA連接酶緩沖液可優(yōu)化連接，可在5分鐘內(nèi)完成。單鏈核酸不是該酶的底物。提供T4 DNA連接酶技術(shù)公告。

應(yīng)用：克隆（鈍端或粘性末端連接）。將接頭或接頭添加到平端DNA。

來源：從大腸桿菌中純化 - 溶原菌NM989。

性能和質(zhì)量測試：內(nèi)切脫氧核糖核酸酶，3'和5'外切脫氧核糖核酸酶測定;結(jié)扎效率測試。

單位定義：一個單位催化在20分鐘內(nèi)將1nmol 32P標記的焦磷酸鹽交換成ATP。在37°C。 （一個單位相當于大約300個粘性末端結(jié)扎單位。）

單位反應(yīng)條件：66mM Tris-HCl（pH 7.6），6.6mM MgCl 2,10mM DTT，66μMATP，3.3μM32P標記的焦磷酸鹽和0.1ml的酶，持續(xù)20分鐘。在37°C。

Invitrogen抗體|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.