T5 Exonuclease

储存缓冲液： 50mM Tris-HCl, 100mM NaCl, 1mM DTT, 0.1mM EDTA, 50% Glycerol, 0.1% Triton X-100, pH 7.5, 25ºC 产品介绍： 阿拉丁生产的T5 Exonuclease，即T5核酸外切酶，是一种按照5'→3'方向降解双链或单链DNA的核酸外切酶。T5 Exonuclease既能从单链或双链DNA 5'末端起始消化，也可以从线性或环状双链DNA的缺口(gap)或缺刻(nick)处起始消化。T5 Exonuclease无法降解超螺旋双链DNA，并且其降解单链DNA的活性可以通过将反应缓冲液中的Mg"2+降低到低于1mM而进行抑制。基于以上特性，T5 Exonuclease常被用于Gibson组装(Gibson Assembly)。Gibson组装是在恒温条件下，有效连接带有多个重叠序列片段的技术，其基本原理可以概括为三步：(1) T5核酸外切酶从DNA片段的5'末端开始消化，产生互补的单链3'末端(overhangs)，促使互补的末端退火；(2) DNA聚合酶填补退火片段的缺口(gaps)；(3) DNA连接酶将组装后的切刻(nicks)处进行连接，最后得到一个完整的双链DNA。 用途： 常用于Gibson组装；降解线性单链、双链DNA或缺刻质粒DNA；从连接的环状双链DNA中去除不完全连接产物；降解线性和缺刻质粒DNA，以获得高纯度的超螺旋质粒DNA；去除碱裂法提取质粒过程中产生的变性质粒DNA；提高小量抽提质粒cDNA文库的转染效率。 活性定义 1、活性单位 (U) 是使双链DNA生成1 nmol酸溶性脱氧核糖核苷酸所需的酶的量，在37ºC条件下，总反应体积为50µl，反应时间为30分钟。 注意事项： T5 Exonuclease是一种对于DNA底物有选择性的核酸外切酶，其对不同的DNA底物显示出不同的反应活性。因此，消化特定的底物时，须注意适当控制好酶量和反应时间。本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。为了您的安全和健康，请穿实验服并戴一次性手套操作。 Storage buffer solution: 50mM Tris-HCl, 100mM NaCl, 1mM DTT, 0.1mM EDTA, 50% Glycerol, 0.1% Triton X-100, pH 7.5, 25ºC Product Introduction: The T5 Exonuclease produced by Aladdin, also known as T5 nuclease, is a nuclease that degrades double stranded or single stranded DNA in the 5 '→ 3' direction. T5 Exonuclease can be digested starting from the 5 'end of single stranded or double stranded DNA, as well as from the gap or nick of linear or circular double stranded DNA. T5 Exonuclease is unable to degrade supercoiled double stranded DNA, and its activity in degrading single stranded DNA can be inhibited by reducing the Mg "2+in the reaction buffer to below 1mM. Based on these characteristics, T5 Exonuclease is commonly used for Gibson Assembly. Gibson assembly is a technique that effectively connects multiple overlapping sequence fragments under constant temperature conditions. Its basic principle can be summarized as three steps: (1) T5 nuclease digests from the 5 'end of the DNA fragment, producing complementary single stranded 3' ends (overhangs) and promoting annealing of the complementary ends; (2) DNA polymerase fills gaps in annealed fragments; (3) DNA ligase connects the assembled nick sites to obtain a complete double stranded DNA. Purpose: Commonly used for Gibson assembly; Degradation of linear single stranded, double stranded DNA or truncated plasmid DNA; Removing incomplete ligation products from connected circular double stranded DNA; Degradation of linear and truncated plasmid DNA to obtain high-purity supercoiled plasmid DNA; Remove the denatured plasmid DNA generated during the process of extracting plasmids using alkaline splitting method; Improve the transfection efficiency of small-scale extraction plasmid cDNA libraries. Definition of Activity 1. The active unit (U) is the amount of enzyme required to generate 1 nmol acid soluble deoxyribonucleotide from double stranded DNA, with a total reaction volume of 50 µ l and a reaction time of 30 minutes at 37 º C. matters needing attention: T5 Exonuclease is a selective nuclease for DNA substrates, exhibiting varying reactivity towards different DNA substrates. Therefore, when digesting specific substrates, it is important to control the enzyme dosage and reaction time appropriately. This product is only for scientific research by professionals and should not be used for clinical diagnosis or treatment, food or medicine, or stored in ordinary residential areas. For your safety and health, please wear lab coats and disposable gloves when operating.

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