NEB/NEBNext Ultrashear™ FFPE DNA Library Prep Kit/24 reactions/E6655S,NEB,,NEB,New England Biolabs,纽英伦,+,,Array.Array.Array蚂蚁淘厂家直采.

产品名称：NEB/NEBNext Ultrashear™ FFPE DNA Library Prep Kit/24 reactions/E6655S

产品编号：E6655S

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

View or download extensive performance data in our Data Supplement.FFPE DNA is often compromised in quality and quantity and is challenging for library prep and sequencing, including in target enrichment workflows that require high amounts of library input. The NEBNext UltraShear FFPE DNA Library Prep Kit addresses these challenges and improves ease of workflow and scalability:

Repairs FFPE-induced damage using the NEBNext FFPE DNA Repair Mix v2
Incorporates specialized enzymatic DNA fragmentation using NEBNext UltraShear
Uses NEBNext Ultra II library prep reagents in a protocol optimized for FFPE DNA
Robustly amplifies libraries using NEBNext MSTC™ FFPE Master Mix
High library yields, sufficient for input into target enrichment workflows
User-friendly workflow with minimal hands-on time and multiple safe stopping points

The NEBNext UltraShear FFPE DNA Library Prep Kit reduces damage from deamination and oxidation due to the fixation, storage, and extraction process for FFPE DNA samples, thereby reducing false positive variant calls. The combination of specialized enzymatic FFPE DNA fragmentation and repair with optimized reagents and protocols provides superior performance with this challenging sample type. For FFPE DNA library prep without enzymatic fragmentation, the NEBNext FFPE DNA Library Prep Kit (NEB #E6650) is available.

Image of workflow — The NEBNext UltraShear FFPE DNA Library Prep Kit has a streamlined workflow with minimal hands-on time across a range of inputs from 5-250 ng. The protocol has been optimized for the user to safely store the reaction after any step in the workflow overnight at -20°C without affecting library yield or quality.

Image of library yields — Libraries were prepared from 5, 50, or 250 ng of normal tissue FFPE DNA ranging in quality from DNA Integrity Number (DIN) 1.8 to 6.8, with the indicated PCR cycles. Libraries were prepared in triplicate for 5 ng and 50 ng input and 1 replicate for 250 ng. Each bar represents the average of triplicates with error bars indicating standard deviation for the 5 and 50 ng inputs. Robust library yields were obtained across sample qualities and input amounts. Most target enrichment workflows require a 200 ng library for hybrid capture input. Sufficient library yield can be obtained using a minimum of 50 ng FFPE DNA with the NEBNext UltraShear FFPE DNA Library Prep Kit.

Image showing library yields — Libraries were prepared in duplicate from 100 ng of low quality, normal tissue FFPE DNA (DIN 1.8) and 9 PCR cycles, using the NEBNext UltraShear™ FFPE DNA Library Prep Kit. Results were compared to other enzymatic fragmentation-based library prep kits that have been validated for use with FFPE samples, using each vendor’s own recommended adaptors (IDT^® xGen^® EZ UNI, Kapa EvoPlus^® Library Prep Kit, QuantaBio^® sparQ DNA Library Prep Kit, and Twist Library Preparation EF 2.0 kit). Libraries were sequenced on the Illumina^® NovaSeq^®6000 (2 x 100 base reads) and downsampled to 5 million paired-end reads. Reads were mapped using Bowtie2 (version 2.3.2.2) to the GRCh38 reference and duplicates were marked using Picard MarkDuplicates (version 1.56.0). Library quality metrics were assessed using Picard Alignment Summary Metrics (version 1.56.0). The level of foldback reads was calculated using Seq_frag_remap (version 0.2). The NEBNext UltraShear FFPE DNA Library Prep Kit improves library quality by reducing the percentage of unmapped, chimeric, non-properly paired, and foldback reads.

Image showing artifact reduction — Libraries were prepared in duplicate from 100 ng of low quality, normal tissue FFPE DNA (DIN 1.8) and 9 PCR cycles, using the NEBNext UltraShear FFPE DNA Library Prep Kit. Results were compared to other enzymatic fragmentation-based library prep kits that have been validated for use with FFPE samples, with each vendor’s own recommended adaptors (IDT xGen EZ UNI, Kapa EvoPlus Library Prep Kit, QuantaBio sparQ DNA Library Prep Kit, and Twist Library Preparation EF 2.0 kit). Libraries were sequenced on the Illumina NovaSeq 6000 (2 x 100 base reads) and downsampled to 5 million paired-end reads. Reads were mapped using Bowtie2 (version 2.3.2.2) to the GRCh38 reference and duplicates marked using Picard MarkDuplicates (version 1.56.0). The average frequency of C→T mutations at each C position (A) and G→T mutations at each G position (B) in Read 1 and 2 was calculated for two technical replicates using Tasmanian (version 1.0.7). C→T mutations arising from cytosine deamination and G→T mutations arising from oxidative damage in low quality FFPE DNA are effectively repaired by the NEBNext FFPE DNA Repair v2 Mix in the NEBNext UltraShear FFPE DNA Library Prep Kit. Other kits show a high level of C→T and G→T artifacts in low quality FFPE DNA due to a lack of DNA damage repair.

Image of hybrid sequencing comps — Libraries were prepared in duplicate from 100 ng of low quality, normal tissue FFPE DNA (DIN 1.8) and 9 PCR cycles, using the NEBNext UltraShear™ FFPE DNA Library Prep Kit. Results were compared to other enzymatic fragmentation-based library prep kits that have been validated for use with FFPE samples, with each vendor’s own recommended adaptors (IDT xGen EZ UNI, Kapa EvoPlus Library Prep Kit, QuantaBio sparQ DNA Library Prep Kit, and Twist Library Preparation EF 2.0 kit). The full library yield was used in singleplex target enrichment with a custom cancer panel (Twist Bioscience^®) and libraries were sequenced on the Illumina NovaSeq 6000 (2 x 100 base reads). 15 million paired-end reads were trimmed with Fastp (version 0.20.0) and mapped with BWA mem (version 0.7.17) to the T2T reference. Duplicates were marked using Picard MarkDuplicates (version 2.20.6) with UMI. Target enrichment quality metrics were assessed using Picard HS Metrics (version 2.18.29). The improved yield, coverage, and fraction of usable reads observed in NEBNext UltraShear FFPE DNA Library Prep Kit whole genome sequencing (WGS) libraries correlates to improved coverage, on-target rate, and coverage uniformity in target enrichment libraries.

This product is related to the following categories:: FFPE DNA,; Next Generation Sequencing Library Preparation

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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