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蚂蚁淘在线 / 品牌中心 / NEB / NEB/NEBNext® Ultra™ II FS DNA Library Prep Kit for Illumina®/24 reactions/E7805S

NEB/NEBNext® Ultra™ II FS DNA Library Prep Kit for Illumina®/24 reactions/E7805S

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货号：E7805S
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The NEBNext® Ultra™ II FS DNA Library Prep Kit for Illumina provides a fragmentation system: a fast and reliable solution for DNA fragmentation and library construction. A new DNA fragmentation reagent is combined with end repair and dA-tailing reagents, allowing these steps to be performed in the same tube, with no clean-ups or sample loss. The same fragmentation protocol is used for any input amount (100 pg–500 ng), or GC content. The kit also includes the NEBNext Ultra II Ligation Master Mix for adaptor ligation, and the NEBNext Ultra II Q5® Master Mix for uniform, high-fidelity library amplification. Please note that adaptors and primers are not included in the kit and are available separately.

For protocols including bisulfite converted DNA, we recommend the NEBNext Ultra II DNA Library Prep Kit for Illumina.Features:

  • Fragmentation, end repair and dA-tailing reagents in a single enzyme mix
  • A single fragmentation protocol, regardless of DNA input amount or GC content
  • Input amounts: 100 pg–500 ng
  • Input DNA can be in water, Tris or TE
  • Workflow: ~ 2.5 hours, with < 15 minutes hands-on time

Download extensive performance datain our technical note.Also available with optional SPRIselect® beadsfor size selection/clean-up steps

For use with NEBNext Multiplex Oligos for Illumina (Unique Dual Index UMI Adaptors DNA Set 1) (NEB #E7395), refer to the Protocols tab for UMI Adaptors-specific guidance.

Figure 1: NEBNext Ultra II FS DNA produces the highest yields, from a range of input amounts
Libraries were prepared from Human NA19240 genomic DNA using the input amounts and numbers of PCR cycles shown. For NEBNext Ultra II FS, a 20-minute fragmentation time was used. For Kapa™HyperPlus libraries, input DNA was cleaned up with 3X beads prior to library construction, as recommended, and a 20-minute fragmentation time was used. Illumina® recommends 50 ng input for Nextera, and not an input range; therefore, only 50 ng was used in this experiment. “Covaris®” libraries were prepared by shearing each input amount in 1X TE Buffer to an insert size of ~200 bp using a Covaris instrument, followed by library construction using the NEBNext Ultra II DNA Library Prep Kit (NEB #E7645). Error bars indicate standard deviation for an average of 3–6 replicates performed by 2 independent users.
Figure 2: NEBNext Ultra II FS DNA provides consistent fragmentation regardless of input amount
Libraries were prepared from Human NA19240 genomic DNA using the input amounts shown. NEBNext Ultra II FS libraries were prepared using a 20-minute fragmentation time. For Kapa™ HyperPlus, input DNA was cleaned up with 3X beads prior to library construction, as recommended, and a 20-minute fragmentation time. Library size was assessed using the Agilent® Bioanalyzer®. Low input (1 ng and below) libraries were loaded on the Bioanalyzer without a dilution. High input libraries were loaded with a 1:5 dilution in 0.1X TE.
Figure 3: NEBNext Ultra II FS DNA provides uniform GC coverage for microbial genomic DNA over a broad range of GC composition
Libraries were prepared using 1 ng of a mix of genomic DNA samples from Haemophilus influenzae, Escherichia coli (K-12 MG1655), Rhodopseudomonas palustris and the library prep kits shown, with 9 PCR cycles for consistency across samples, and sequenced on an Illumina MiSeq®. NEBNext Ultra II FS libraries were prepared using a 20-minute fragmentation time. For Kapa HyperPlus libraries, input DNA was cleaned up with 3X beads prior to library construction, as recommended, followed by a 25-minute fragmentation time. “Covaris” libraries were prepared by shearing 1 ng of DNA in 1X TE Buffer to an insert size of ~200 bp using a Covaris instrument, followed by library construction using the NEBNext Ultra II DNA Library Prep Kit (NEB #E7645). Reads were mapped using Bowtie 2.2.4 and GC coverage information was calculated using Picard’s CollectGCBiasMetrics (v1.117). Expected normalized coverage of 1.0 is indicated by the horizontal grey line, the number of 100 bp regions at each GC% is indicated by the vertical grey bars, and the colored lines represent the normalized coverage for each library.
Figure 4: ULTRA II FS produced 10-15x more DNA library compared with other methods -Data from Peter Ellis of the Wellcome Trust Sanger Institute
Human genomic DNA was subjected to DNA library construction using an existing DNA library construction workflow (CURRENT), NEB Ultra II or NEB Ultra II FS. Adapter-Ligated libraries were amplified by PCR (4-12 cycles), purified and quantitated using the Agilent Bioanalyzer platform. Values obtained were used to normalize DNA library yield from 12 cycles of PCR.

View additional data presented at AGBT by Peter Ellis, Senior Staff Scientist at the Wellcome Trust Sanger Institute.

This product is related to the following categories:
DNA Fragmentation & RNA Fragmentation,
DNA Library Prep for Illumina,
Automation for NEBNext® NGS Library Prep
New England Biolabs（NEB）公司       NEB公司——三十多年的卓越品质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。重组酶NEB公司对酶的生产与其基础科研不能分开，通过对限制/修饰系统的克隆和过量表达方面的研究，使我们能够大大降低成本，改善产品质量。NEB已经成功克隆了180多种内切酶，其中大多是完全克隆，少数是部分克隆。目前，NEB可供应240多种内切酶，其中180多种可以重组酶形式提供，同时还有大量的应用途广泛的重组聚合酶和重组修饰酶。质量与客户服务NEB公司凭借着严格的质检程序、深入的基础科研以及不断开发的研发项目，承诺为全球科技人员提供高纯度的科研产品。直接与NEB总部联系或与NEB的国际网点联系，即可体验到NEB的个性化客户服务。公司内负责生产以及负责进行质量监控的科学家（产品负责人）也就是技术支持人员。他们为客户解答有关限制核酸内切酶、甲基化酶、以及其他DNA修饰酶、蛋白质修饰酶方面的问题。同时，公司的有机合成部门可提供linkers、引物、adaptors、探针以及寡核苷酸合成等方面的信息。研究人员也可为客户提供技术支持服务，这些研究人员在DNA测序、甲基化、克隆、过量表达、发酵、蛋白质纯化以及蛋白质分析方面都有很深造诣。基础科研NEB在分子生物学和寄生虫学方面的基础研究由公司内部的资深科学家负责。这些科学家在他们各自的领域都卓有成绩，经常在业内著名杂志上发表文献，指导博士后工作、为大学生提供暑期实习机会，而且经常受邀去当地学校进行演讲。NEB鼓励公司与外界合作，每周学术探讨会为其他科学家们提供了交流和展示的平台。环保政策NEB公司在资助科研项目时最基本原则之一就是：一切工作都应符合保护生态环境。公司产品生产、分析以及运输过程既要满足产品的稳定性，也要符合我们的环保政策。20年来，NEB公司最引人注目的一项环保政策是：回收运输泡沫盒，虽然这项工作首创于美国，但是，目前加拿大、德国以及英国的分公司也同样实施了。公司还在内部开展多方位的回收工作，经常用再生纸张印刷市场宣传资料。这些小细节不会影响产品质量，但是对环境的保护却是显而易见。公司职责除环保政策外，通过基金会，NEB还竭力为改善当地社区及全人类尽微薄之力。NEB基金会创立于1982年，属私人性质，其宗旨是支持发展中国家的环保、教育、健康及艺术事业。此外，NEB捐款委员会也经常向当地社区进行捐款活动。