NEB/EnGen® Mutation Detection Kit/25 reactions/E3321S,NEB,,NEB,New England Biolabs,纽英伦,+,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：NEB/EnGen® Mutation Detection Kit/25 reactions/E3321S

产品编号：E3321S

市场价：¥0.00

场地：美国(厂家直采)

产品分类：分子类>核酸酶类>其他>

联系QQ：1570468124

电话号码：4000-520-616

邮箱： info@ebiomall.com

美元价：待定

品牌： NEB

公司：New England Biolabs

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商品介绍

The EnGen Mutation Detection Kit provides reagents for detection of on-target genome editing events. In the first step, targeted regions from cells whose genomes were targeted (i.e. CRISPR/Cas9, TALENs, Zinc-finger Nucleases) are amplified using Q5 Hot Start High-Fidelity 2X Master Mix. Upon denaturation and re-annealing, heteroduplexes are formed when mutations from insertions and deletions (indels) are present in the amplicon pool. In the second step, annealed PCR products are digested with EnGen T7 Endonuclease I, a structure-specific enzyme that will recognize mismatches larger than 1 base. Both strands of the DNA are cut when a mismatch is present, which results in the formation of smaller fragments. Analysis of the resulting fragments provides an estimate of the efficiency of the genome editing experiments. The EnGen Mutation Detection Kit includes a Control Template and Primer Mix that can be used as a control for the PCR reaction and T7 Endonuclease I digestion. The Control Template and Primer Mix provided contains two plasmids and primers that when amplified, denatured and re-annealed will form heteroduplexes that contain a 10-base insertion. This structure is a substrate for T7 Endonuclease I. The digestion of the 600 bp heteroduplex containing amplicon yields products of 200 bp and 400 bp. 600 bp parental homoduplexes are uncleaved, and are easily distinguished from cleaved heteroduplexes when separated and visualized by agarose gel electrophoresis or fragment analysis instrument.The protocol has been optimized so that PCR products generated by the Q5 Hot Start High-Fidelity 2X Master Mix can be introduced directly into the T7 Endonuclease I digestion without the need for purification. Digestion of the heteroduplex is complete in only 15 minutes, and Proteinase K is included to stop the reaction efficiently. Additional Q5 Hot Start High-Fidelity 2X Master Mix is also included to allow for optimization of target site amplification before digestion.Figure 1: Workflow for Mutation Detection Kit

Genomic DNA is amplified with primers bracketing the modified locus. PCR products are then denatured and re-annealed yielding three classes of possible structures. Duplexes containing a mismatch greater than one base are digested by T7 Endonuclease I. The DNA is then electrophoretically separated and fragment analysis is used to estimate targeting efficiency.

This product is related to the following categories:: Mutation Detection Reagents and Kits,; Genome Editing Products,; DNA Repair Enzymes and Structure-specific Endonucleases Products

This product can be used in the following applications:: Genome Editing Applications

品牌介绍

NEB公司成立于二十世纪七十年代中期，拥有众多经验丰富的科学家，是生产生命科学试剂的领导者。目前，NEB为基因组研究提供最齐全的重组酶和天然酶，并且公司业务范围已延伸至蛋白质组学和药品开发领域。回顾三十余年来的历程，NEB公司作为先驱公司之一，为促进生物科技工业的发展做出了巨大的贡献。NEB美国总部乔迁新址后拥有最尖端的设备，有一座现代化的发酵中心及设备齐全的实验室，这些实验室主要用于产品生产、质量监控、产品开发和基础科研之用。作为首批以商业规模生产限制性内切酶的公司之一，NEB一直专注于内切酶的研究，并保持业内领先水平。NEB公司一贯坚持以科学为本的原则，公司生产的试剂因其高质量、高性价比享誉世界。NEB（北京）有限公司 New England Biolabs (Beijing) LTD. NEB（北京）有限公司New England Biolabs (Beijing) LTD. 为美国New England Biolabs, Inc.在华投资兴建的独资子公司，负责处理其在中国内地及香港地区的全部业务，同时承担部分新产品的研发工作。纽英伦生物技术（北京）有限公司成立于2001年11月，为中国生命科学研究者提供一流的产品和专业的技术服务，促进国内外生命科学学术交流，与中国科研工作者一起将中国推入到二十一世纪生命科学的前沿中去。

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