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主要产品:NEB、内切酶、基因表达、RNAi DNA修饰酶、各种载体、 Marker 、引物、测序 等等
℡ 4000-520-616
℡ 4000-520-616
NEB/NEBNext® FFPE DNA Library Prep Kit/24 reactions/E6650S
产品编号:E6650S
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:
NEB/NEBNext® FFPE DNA Library Prep Kit/24 reactions/E6650S
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 a challenging sample type for library prep and sequencing. The NEBNext FFPE DNA Library Prep Kit addresses these challenges in multiple ways:

  • Repairs FFPE-induced damage using the NEBNext FFPE DNA Repair Mix v2
  • 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 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. For further performance improvement, the NEBNext UltraShear FFPE DNA Library Prep Kit (NEB #E6655) also incorporates specialized enzymatic DNA fragmentation.

FIGURE 1: NEBNext FFPE DNA Library Prep Kit workflow Image of Workflow
The NEBNext FFPE DNA Library Prep kit has a streamlined workflow with minimal hands-on time. 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.
FIGURE 2: The NEBNext FFPE DNA Library Prep Kit enables robust library preparation from a range of sample inputs and quality Image of library yield
Libraries were prepared from 5, 50 or 250 ng of Covaris®-sheared normal tissue FFPE DNA ranging in quality from DNA Integrity Number (DIN) 1.8 to 6.8 with the indicated PCR cycles. Libraries were made in triplicate for 5 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 200 ng of library for hybrid capture input, and sufficient library yield can be obtained using a minimum of 50 ng FFPE DNA with the NEBNext FFPE DNA Library Prep Kit.
FIGURE 3: NEBNext FFPE DNA Library Prep Kit reduces damage-derived sequencing artifacts Image ofartifact reduction graph
Libraries were prepared from 50 ng of Covaris-sheared normal tissue FFPE DNA of either low (DIN 1.8) or high (DIN 6.8) quality using the NEBNext FFPE DNA Library Prep Kit and other library prep kits as shown. Libraries were sequenced on the Illumina® NextSeq® 500 (2 x 76 bases). Libraries were downsampled to 600,000 paired-end reads, mapped using bowtie2 (version 2.3.2.2) to the GrCh38 reference, and duplicates marked using Picard MarkDuplicate (version 1.56.0). The average frequency of C→T mutations at each C position in Read 2 was calculated for two technical replicates using Tasmanian (version 1.0.7). C→T mutations arising from cytosine deamination damage in low quality FFPE DNA are effectively repaired by the NEBNext FFPE DNA Repair v2 mix included in the NEBNext FFPE DNA Library Prep Kit. Other kits show a high level of C→T artifacts in low quality FFPE DNA (DIN 1.8) due to a lack of DNA damage repair.
FIGURE 4: The NEBNext FFPE DNA Library Prep Kit detects expected variants in formalin-compromised reference standard DNA IMage of charts on expected variants
Libraries were prepared in triplicate from 50, 100, or 250 ng of Covaris-sheared formalin-compromised reference standard DNA (severe) (Horizon Discovery HD803) using either the NEBNext FFPE DNA Library Prep Kit or NEBNext Ultra™ II DNA Library Prep Kit. The full library yield was used in a target enrichment workflow with a custom cancer panel (Twist Bioscience®) as a 6-plex capture reaction. Libraries were sequenced on the Illumina NovaSeq® 6000 (2 x 100 bases). All fastq files were downsampled to 22 million paired-end reads, mapped using BWA mem (version 0.7.17) to the T2T reference, and duplicates marked using Picard MarkDuplicates (version 1.56.0). Variant allele frequencies (VAF) were calculated from Mpileup using Samtools (version 1.16.1) and plotted against the expected VAF in the Horizon reference DNA. A variant with a frequency of 0.88% was missed at 50 ng input for 1/3 replicates, but all variants were detected in all replicates at 100 and 250 ng input. The NEBNext FFPE DNA Library Prep kit improves the correlation of expected to observed VAF in low input libraries (50 ng) compared to standard library prep (NEBNext Ultra II DNA), indicating the benefit of the yield and coverage obtained with the NEBNext FFPE DNA Library Prep Kit including repair with NEBNext FFPE DNA Repair v2.
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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