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Monarch® gDNA Nuclei Prep & Lysis Buffer Pack |NEB酶试剂 New England Biolabs

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上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

The Monarch gDNA Nuclei Prep & Lysis Buffer Pack contains two lysis buffers used in the Monarch HMW DNA Extraction Kit for Cells & Blood (NEB #T3050). The Nuclei Prep Buffer is a mild lysis buffer used first to lyse the cell membrane and release the contents of the cytoplasm while the nuclei are kept intact. It is used pre-mixed with the RNase A enzyme included in the kit. After the RNA from the cytoplasm is digested by RNase A, the Nuclei Lysis Buffer, pre-mixed with the included Proteinase K, is used to lyse the nuclei and release the HMW genomic DNA. This two-step lysis ensures that RNA in the cytoplasm is effectively degraded before the viscous genomic DNA from the nuclei is released. 20 ml of each buffer is included in this buffer pack. The Nuclei Prep Buffer is used chilled to 4°C, and it can be stored at 4°C for convenience if desired.

产品类别:
Genomic DNA Extraction & Purification Products,
Nucleic Acid Purification Products

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • T3054L     25    
        Monarch® gDNA Nuclei Lysis Buffer T3053-1 25 2 x 10 ml Not Applicable
        Monarch® gDNA Nuclei Prep Buffer T3052-1 25 2 x 10 ml Not Applicable

  • 相关产品

    相关产品

    • Monarch 高分子量 DNA 提取试剂盒（细胞和血液）
    • T3060 Monarch HMW DNA Extraction Kit for Tissue
    • Monarch RBC 裂解液
    • T3056 Monarch gDNA Elution Buffer II
    • Monarch DNA 捕获珠
    • T3056 Monarch gDNA Elution Buffer II
    • Monarch 捕获珠分离管
    • Monarch® 收集管 II
    • p8107-proteinase-k-molecular-biology-grade
    • Monarch 2 ml 离心管
    • T3015 Monarch gDNA Wash Buffer

操作说明、说明书 & 用法

  • 操作说明

    1. Download Quick Protocol Card for Cells
    2. Download Quick Protocol Card for Blood
    3. Protocol for High Molecular Weight DNA (HMW DNA) Extraction from Cells (NEB #T3050)
    4. Protocol for High Molecular Weight DNA (HMW DNA) Extraction from Blood (NEB #T3050)
    5. Protocol for UHMW DNA Cleanup in the Oxford Nanopore Technologies® UL Library Prep Workflow

FAQs & 问题解决指南

  • 问题解决指南

    • Troubleshooting Guide for Monarch HMW DNA Extraction Kits (NEB #T3050 and #T3060)

Next® FFPE DNA Library Prep Kit |NEB酶试剂 New England Biolabs

发表于由

上海金畔生物科技有限公司代理New England Biolabs（NEB）酶试剂全线产品，欢迎访问官网了解更多产品信息和订购。

产品信息

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

Next® FFPE DNA Library Prep Kit |

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

Next® FFPE DNA Library Prep Kit |

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

Next® FFPE DNA Library Prep Kit |    

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

Next® FFPE DNA Library Prep Kit |

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.

产品类别:
FFPE DNA Products,
Next Generation Sequencing Library Preparation Products

  • 试剂盒组成

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • E6650S     Multi-temperature    
        NEBNext® FFPE DNA Library Prep Kit E6650-2 -20    
        NEBNext® FFPE DNA Repair Mix v2 E7361AVIAL -20 1 x 0.048 ml Not Applicable
        NEBNext® Thermolabile Proteinase K E7362AVIAL -20 1 x 0.048 ml Not Applicable
        NEBNext® FFPE DNA Repair Buffer v2 E7363AVIAL -20 1 x 0.168 ml Not Applicable
        NEBNext® Ultra II End Prep Enzyme Mix E7646AVIAL -20 1 x 0.072 ml Not Applicable
        NEBNext® Ultra™ II Ligation Master Mix E7648AVIAL -20 1 x 0.72 ml Not Applicable
        NEBNext® Ligation Enhancer E7374AVIAL -20 1 x 0.024 ml Not Applicable
        NEBNext® MSTC FFPE Master Mix E6651AVIAL -20 1 x 0.6 ml 2 X
        NEBNext® Sample Purification Beads E6652S 25    
        NEBNext® Sample Purification Beads E6652AVIAL 25 1 x 4.3 ml Not Applicable
    • E6650L     Multi-temperature    
        NEBNext® FFPE DNA Library Prep Kit E6650-3 -20    
        NEBNext® FFPE DNA Repair Mix v2 E7361AAVIAL -20 1 x 0.192 ml Not Applicable
        NEBNext® Thermolabile Proteinase K E7362AAVIAL -20 1 x 0.192 ml Not Applicable
        NEBNext® FFPE DNA Repair Buffer v2 E7363AAVIAL -20 1 x 0.672 ml Not Applicable
        NEBNext® Ultra II End Prep Enzyme Mix E7646AAVIAL -20 1 x 0.288 ml Not Applicable
        NEBNext® Ultra™ II Ligation Master Mix E7648AAVIAL -20 3 x 0.96 ml Not Applicable
        NEBNext® Ligation Enhancer E7374AAVIAL -20 1 x 0.096 ml Not Applicable
        NEBNext® MSTC FFPE Master Mix E6651AAVIAL -20 2 x 1.2 ml 2 X
        NEBNext® Sample Purification Beads E6652L 25    
        NEBNext® Sample Purification Beads E6652AAVIAL 25 1 x 17 ml Not Applicable

  • 特性和用法

    需要但不提供的材料

    • 80% Ethanol
    • Nuclease-free Water
    • 0.1X TE (1 mM Tris-HCl, pH 8.0, 0.1 mM EDTA)
    • DNase-, RNase-free PCR strip tubes
    • NEBNext Multiplex Oligos for Illumina® (www.neb.com/oligos)
    • Magnetic rack/stand (NEB #S1515S, Alpaqua® cat. #A001322, or equivalent)
    • Thermal cycler
    • Agilent® Bioanalyzer® or TapeStation® and associated reagents and consumables
    • Adaptor Dilution Buffer NEB #B1430S or NEBNext Unique Dual Index UMI Adaptor Dilution Buffer supplied with NEBNext Unique Dual Index UMI Adaptor DNA Sets (NEB #E7395/E7874/E7876/E7878)

  • 相关产品

    相关产品

    • e6655nebnext-ultrashear-ffpe-dna-library-pep-kit
    • NEBNext® 多样本接头引物试剂盒 1（96 种 Unique 双端 Index 引物）
    • NEBNext® 多样本接头引物试剂盒 2（96 种 Unique 双端 Index 引物）
    • NEBNext® Multiplex Oligos for Illumina® (96 Unique Dual Index Primer Pairs Set 3)

操作说明、说明书 & 用法

  • 操作说明

    1. Where can I find guidelines and protocols for use of the NEBNext FFPE DNA Library Prep Kit (NEB #E6650)?

  • 说明书

    产品说明书包含产品使用的详细信息、产品配方和质控分析。

    • manualE6650

工具 & 资源

  • 选择指南

    • NEBNext® Multiplex Oligos Selection Chart

FAQs & 问题解决指南

  • FAQs

    1. What sample types can I use with the NEBNext FFPE DNA Library Prep Kit?
    2. How much starting material is required for the NEBNext FFPE DNA Library Prep Kit?
    3. Is size selection recommended?
    4. What are the safe stopping points in this protocol?
    5. What input amount (nanograms) is recommended for standard versus target enrichment library prep with FFPE DNA samples?
    6. Can I use this NEBNext kit with adaptors and primers from vendors other than NEB?
    7. What do I do if I see a precipitate in the NEBNext FFPE DNA Repair Buffer v2?
    8. Will treating my DNA with the FFPE DNA Repair Mix v2 as part of the NEBNext FFPE DNA Library Prep Kit hurt my downstream reaction?
    9. Which NEBNext Multiplex Oligos can be used with the NEBNext FFPE DNA Library Prep Kit?
    10. Does the FFPE DNA Repair v2 Mix repair DNA-protein crosslinks?
    11. Does the FFPE DNA Repair v2 Mix fix blocked 3′ ends?
    12. Can the FFPE DNA Repair v2 Mix repair damage in both single- and double-stranded DNA? Or, does it require double stranded DNA as a template?
    13. If I had a DNA template with mutation sites (i.e. 8-oxoguanine or deaminated cytosines) that are directly adjacent to each other on opposite strands would treatment with the FFPE DNA Repair v2 Mix cause a double strand nick/break?
    14. What gap lengths can be repaired with the FFPE DNA Repair v2 Mix?

Plasmid Mini Prep Kit 高纯质粒小提试剂盒

发表于由

Plasmid Mini Prep Kit

高纯质粒小提试剂盒

品牌：Nippon Gene
CAS No.：
储存条件：-20℃
纯度：–
产品编号

(生产商编号)

 等级 规格 运输包装 零售价(RMB) 库存情况 参考值

311-01521

 – 60 tests –

Plasmid Mini Prep Kit 高纯质粒小提试剂盒

 – –


* 干冰运输、大包装及大批量的产品需酌情添加运输费用


* 零售价、促销产品折扣、运输费用、库存情况、产品及包装规格可能因各种原因有所变动，恕不另行通知，确切详情请联系上海金畔生物科技有限公司。

Transparent Skeletal Specimen Prep Kit 透明骨架样品制备试剂盒

发表于由

Transparent Skeletal Specimen Prep Kit

透明骨架样品制备试剂盒

品牌：FUJIFILM Wako
CAS No.：
储存条件：2-10℃
纯度：–
产品编号

(生产商编号)

 等级 规格 运输包装 零售价(RMB) 库存情况 参考值

295-77301

 for Education 1Kit（4只青鳉鱼用） – 咨询 – –


* 干冰运输、大包装及大批量的产品需酌情添加运输费用


* 零售价、促销产品折扣、运输费用、库存情况、产品及包装规格可能因各种原因有所变动，恕不另行通知，确切详情请联系上海金畔生物科技有限公司。