NEB/NEBridge® Ligase Master Mix/50 reactions/M1100S,NEB,,NEB,New England Biolabs,纽英伦,+,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：NEB/NEBridge® Ligase Master Mix/50 reactions/M1100S

产品编号：M1100S

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

公司分类：

商品介绍

NEBridge^® Ligase Master Mix is a 3X master mix for Golden Gate Assembly. Designed for use with NEB Type IIS restriction enzymes, this master mix contains T4 DNA Ligase in an optimized reaction buffer with a proprietary ligation enhancer. Users need only choose their preferred NEB Type IIS restriction enzyme and add DNA substrates to be assembled. Low complexity single fragment insertions, as well as moderate complexity (3–6 fragment) and high complexity (7–25+ fragment) assemblies, are all supported with this optimized reagent and accompanying protocols.

A. Schematic of 25- fragment lac cassette assembly with NEBridge Ligase Master MixB: *30 µl reactions containing 24 inserts and pGGAselect plasmid were set up using *BbsI-HF (NEB #R3539M)*.*Left plate shows results with 10 µl NEBridge Ligase Master Mix. Right plate shows results with 3 µl of 10X T4 DNA Ligase Buffer (NEB #B0202) and 1 µl of T4 DNA Ligase (NEB #M0202T/M). 1 µl of BbsI-HF was used for both reactions, as well as 0.05 pmol of each 24 insert DNAs and pGGAselect. Reactions proceeded for 60 cycles at 37°C for 5 min and 16°C for 5 min, with an end soak at 60°C for 5 min. 2 µl of each reaction was transformed in to 50 µl of T7 Express Competent E.coli(NEB #C2566). 50 µl of outgrowth was plated onto LB plates (supplemented with 1 mg/ml dextrose, 1 mg/ml MgCl₂, 30 µg/ml chloramphenicol, 200 µM IPTG and 80 µg/ml X-gal) and incubated at 37°C for 18 hrs, and then stored at 4°C for 4 hrs before scoring colony color phenotype.C: The total transformants and percentage of correct assemblies (blue colonies) were reported as the average result of three replicates with the standard deviation from the mean. The reaction with NEBridge Ligase Master Mix generated 2.2 ±0.2X10⁶ correctly assembled blue colonies per µg vector DNA with 94.3 ±1% fidelity, while the reaction with T4 DNA Ligase Buffer generated 8.3 ±2.1X10⁵ correctly assembled blue colonies per µg vector DNA with 69.8 ± 10.7% fidelity.

ReferenceV. Potapov et al., Comprehensive Profiling of Four Base Overhang Ligation Fidelity by T4 DNA Ligase and Application to DNA Assembly. ACS Synth Biol 7, 2665–2674 (2018).

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

This product can be used in the following applications:: High-throughput cloning and automation solutions,; NEBridge® Golden Gate 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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