NEB/Gibson Assembly® Cloning Kit/10 reactions/E5510S,NEB,,NEB,New England Biolabs,纽英伦,+,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：NEB/Gibson Assembly® Cloning Kit/10 reactions/E5510S

产品编号：E5510S

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

Gibson Assembly was developed by Dr. Daniel Gibson and his colleagues atthe J. Craig Venter Institute and licensed to NEB by Synthetic Genomics, Inc. Itallows for successful assembly of multiple DNA fragments, regardless of fragmentlength or end compatibility. It has been rapidly adopted by the syntheticbiology community due to its ease-of-use, flexibility and suitability for large DNAconstructs.Gibson Assembly efficiently joins multiple overlapping DNA fragments in asingle-tube isothermal reaction (1,2). The Gibson Assembly Master Mix includesthree different enzymatic activities that perform in a single buffer:

The exonuclease creates single-stranded 3´ overhangs that facilitate theannealing of fragments that share complementarity at one end (overlap region).
The proprietary DNA polymerase fills in gaps within each annealed fragment.
The DNA ligase seals nicks in the assembled DNA.

The end result is a double-stranded fully sealed DNA molecule that can serveas template for PCR, RCA or a variety of other molecular biology applications,including direct transformation. The method has been successfully used by Gibson’sgroup and others to assemble oligonucleotides, DNA with varied overlaps(15–80 bp) and fragments hundreds of kilobases long (1–2).To help select the best DNA assembly method for your needs, please use our Synthetic Biology/DNA Assembly Selection Chart.For help designing primers, please view our primer design video.

Overview of the Gibson Assembly Cloning Method

Specification:

10 μl of 2X Gibson Assembly Master Mix was incubated with 6 fragments(5 fragments of 400 bp and one of 2,780 bp, with 40 bp overlap, 0.05 pmoleach) in a final volume of 20 μl at 50°C for 60 minutes. NEB 5-alpha CompetentE. coli (NEB #C2987) were transformed with 2 μl of the master mix/fragmentmixture using the transformation protocol on page 12. Greater than 100white colonies were observed when 1/10 of the outgrowth was spread on anampicillin plate with IPTG/Xgal and incubated overnight.Overview of Gibson Assembly Cloning Kit Protocol:

Design primers to amplify fragments (and/or vector) with appropriateoverlaps
PCR amplify fragments using a high-fidelity DNA polymerase.
Prepare linearized vector by PCR amplification using a high-fidelity DNA polymerase or by restriction digestion.
Confirm and determine concentration of fragments and linearized vector using agarose gel electrophoresis, a NanoDrop™ instrument or other method.
Add fragments and linearized vector to Gibson Assembly Master Mixand incubate at 50°C for 15 minutes to 1 hour, depending on number offragments being assembled.
Transform into NEB 5-alpha Competent E. coli (provided) or use directly inother applications.

This product is related to the following categories:: DNA Assembly, Cloning and Mutagenesis Kits Products

This product can be used in the following applications:: Gibson Assembly®

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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