StrataClone PCR 克隆试剂盒-分析测试百科网

StrataClone PCR 克隆试剂盒

面议

品牌：安捷伦
厂商性质：生产商
产地类别：进口
产地：美国
货期：现货
服务地域：全国
规格：3
近期咨询：1

Agilent StrataClone PCR 克隆试剂盒可在室温下高效克隆 PCR 产物，仅需 5 分钟。该试剂盒利用 DNA 拓扑异构酶 I 的 DNA 重接活性和 Cre 重组酶的 DNA 重组活性，产生 > 95％插入片段的

400-6699-117 转1000

货号	规格	操作
240205	StrataClone PCR Cloning Kit	询底价
240207	StrataClone Blunt PCR Cloning Kit	询底价
240218	StrataClone Ultra Blunt PCR Cloning Kit	询底价

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耗材描述

Agilent StrataClone PCR 克隆试剂盒可在室温下高效克隆 PCR 产物，仅需 5 分钟。该试剂盒利用 DNA 拓扑异构酶 I 的 DNA 重接活性和 Cre 重组酶的 DNA 重组活性，产生 > 95％插入片段的阳性克隆。该试剂盒可用于平端和 UA 克隆，可以克隆长短扩增子。

此外，我们还提供 StrataClone Ultra PCR 克隆系统。该试剂盒包含 PfuUltra II 融合热启动 DNA 聚合酶。StrataClone Ultra 系统将 PCR 扩增的最高准确度与简单、可靠且价格合理的基于拓扑异构酶的克隆相结合。

在室温下克隆 PCR 产物仅需 5 分钟。
结合 DNA 拓扑异构酶 I 的 DNA 重接活性和 Cre 重组酶的 DNA 重组活性。
可用于平端和 UA PCR 克隆。
效率高，产生 > 95％插入片段的阳性克隆。
使用同一款试剂盒克隆长和短扩增子 (500 bp–9.2 Kb)。
包括卡那霉素和氨苄青霉素抗性标记。

该产品可根据您的需求进行定制。

工作原理

Overview of StrataClone PCR Cloning Technology

This technology exploits the activities of topoisomerase I from Vaccinia virus and Cre recombinase from bacteriophage P1. In vivo, DNA topoisomerase I assists in DNA replication by relaxing and rejoining DNA strands. Topoisomerase I cleaves the phosphodiester backbone of DNA strands after the sequence 5´-CCCTT. it forms a covalent DNA–enzyme intermediate that conserves bond energy for religating the cleaved DNA back to the original strand. Once the covalent DNA–enzyme intermediate is formed, the religation reaction can also occur with a heterologous DNA acceptor.

Two DNA arms

The Cre recombinase enzyme catalyzes recombination between two loxP recognition sequences.

The StrataClone PCR cloning vector mix contains two DNA arms, each charged with topoisomerase I on one end and containing a loxP recognition sequence on the other end. The topoisomerase-charged ends have a modified uridine (U*) overhang. Taq-amplified PCR products, which contain 3´-adenosine overhangs, are efficiently ligated to these vector arms in a 5-minute ligation reaction, through A-U* base-pairing followed by topoisomerase I-mediated strand ligation.

StrataClone PCR cloning vector pSC-A-amp/kan

The resulting linear molecule is transformed into a competent cell line, engineered to transiently express Cre recombinase. Cre-mediated recombination between vector loxP sites creates a circular DNA molecule that is proficient for replication in cells growing on media containing ampicillin or kanamycin. Resulting pSC-A-amp/kan vector product includes a lacZ´ a-complementation cassette for blue-white screening. The circular map shows the topoisomerase I-mediated ligation product of the supplied vector arms to a PCR product of interest followed by Cre-mediated recombination.

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