Illumina DNA Prep with Enrichment

快速、集成的工作流程,为广泛的靶向测序应用提供可直接用于测序的DNA文库。

约6.5小时

分析时间

约2小时

手动操作时间

10 -1000 ng高质量基因组DNA或50-1000 ng FFPE DNA。(对于血液和唾液样本,请参阅参考指南)。

起始量

详情请参见规格表

概述

Illumina DNA Prep with Enrichment,原名为Nextera Flex for Enrichment,是Illumina文库制备产品系列中最快、最灵活的DNA靶向测序解决方案。

  • 仅需6.5小时即可完成文库制备、富集和富集后扩增

  • 通过血液和唾液的集成实验方案提高文库制备效率

  • 通过高度均一的插入片段大小获得一致的文库产量

快速、用户友好的工作流程

通过内置定量和文库归一化,磁珠上酶切法片段化化学技术与简化的单一杂交实验方案相结合,将手动操作时间缩短至约2小时。

样本起始量灵活性

兼容多种DNA起始量和样本类型,包括血液、唾液、基因组DNA和福尔马林固定石蜡包埋(FFPE)组织。

经过优化的性能

磁珠上酶切法片段化可最大限度减少偏差和出错机会,从而在所有Illumina测序系统中获得高度可重现的数据。


Illumina DNA Prep产品系列

查找快速、优化的测序文库制备解决方案,适用于各种应用。


性能参数


所需产品

Illumina DNA Prep and Enrichment试剂盒需要使用Illumina DNA/RNA UD Indexes Set。建议使用双标签。 

Panel和探针是可选的,请单独订购。

Flex Lysis Kit是对血液DNA提取的补充。

鉴定FFPE样本需要Infinium FFPE QC Kit。

使用SciGene Hybex Microheating系统时,Infinium MIDI Heatblock Insert是必需的。 

Illumina Free Adapter Blocking Reagent(可选)用于降低标签跳跃水平。

Illumina纯化磁珠用于DNA纯化和/或根据磁珠与样本的比例选择DNA片段的大小。

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应用

Illumina DNA Prep with Enrichment结合了多种DNA起始量和样本类型的多功能、简单、快速的文库制备和富集,支持广泛的靶向富集应用,包括定制panel、固定panel和全外显子组panel。

工作流程示例



项目建议

仪器 Recommended number of samples Read length
iSeq 100 System

TruSight Cancer: 4 samples per run (based on 280× mean targeted coverage; 99% target coverage at 20×)
TruSight Cardio: 4 samples per run (based on 170× mean targeted coverage; 100% target coverage at 20×)

2×100 bp

2×100 bp

MiniSeq System

TruSight Cancer (samples per run): mid output: 8, high output: 25 (based on 280× mean targeted coverage; 99% target coverage at 20×)
TruSight Cardio (samples per run): mid output: 8, high output: 25 (based on 170× mean targeted coverage; 100% target coverage at 20×)

2×100 bp

2×100 bp

MiSeq System

TruSight Cancer (samples per run): v2 reagents: 15, v3 reagents: 25 (based on 280× mean targeted coverage; 99% target coverage at 20×)
TruSight Cardio (samples per run): v2 reagents: 15, v3 reagents: 25 (based on 170× mean targeted coverage; 100% target coverage at 20×)
TruSight One (samples per run): v2 reagents: 1, v3 reagents: 3 (based on 120× mean targeted coverage; 95% target coverage at 20×)

2×100 bp

2×100 bp

2 x 100 bp(最多2 x 150 bp)

NextSeq 550 System

Exome (samples per run): mid output: 5, high output: 16 (based on 50× mean targeted coverage; 90% target coverage at 20×)

TruSight One (samples per run): mid output: 12, high output: 36 (based on 120× mean targeted coverage; 95% target coverage at 20×)
TruSight One Expanded (samples per run): mid output: 7, high output: 24 (based on 130× mean targeted coverage; 97% target coverage at 20×)

2×100 bp
2 x 100 bp(最多2 x 150 bp)
2 x 100 bp(最多2 x 150 bp)

NovaSeq 6000 System

Exome (samples per run, by flow cell type): S1: 128, S2: 164, S4: 400 (based on 50× mean targeted coverage; 90% target coverage at 20×; S4 limited by available indexes)

2×100 bp


相关应用和方法

比较

选择工具:

Illumina磁珠上酶切法片段化化学技术

由eBLT介导的均匀片段化反应,加上单一杂交反应,可实现快速、灵活的工作流程。

富集panel兼容性

Illumina DNA Prep with Enrichment与各种Illumina富集panel兼容,并针对低通量、中通量和高通量系统进行了优化。

高覆盖度均一性和加垫read富集

Illumina DNA Prep with Enrichment为定制、固定和外显子组panel提供了高覆盖度均一性和靶向填充read富集。

准确的变异检出

Illumina DNA Prep with Enrichment为(A、B)细胞系FFPE对照人类参考DNA样本和(C、D)真实世界FFPE样本提供低丰度体细胞变异检出,观察到的变异频率与直系同源测序检测的频率显著相关。

文库制备 (4)

Illumina® DNA Prep with Enrichment, (S) Tagmentation (96 Samples)

20025524

包括用于制备和富集96个文库的试剂(8次12重富集反应)。单独购买富集探针panel、Illumina纯化磁珠和标签接头。

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Illumina® DNA Prep with Enrichment, (S) Tagmentation (16 Samples)

20025523

包含用于制备和富集16个文库的试剂(16,1重富集反应)。单独购买富集探针panel、Illumina纯化磁珠和标签接头。

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Illumina® DNA Prep, (S) Tagmentation (96 Samples)

20025520

包含用于制备96个文库的试剂。不包含富集试剂。单独购买富集探针panel、Illumina纯化磁珠和标签接头。富集试剂只能与文库制备试剂(Illumina DNA Prep with Enrichment, (S) Tagmentation(96样本))一起订购。

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Illumina® DNA Prep, (S) Tagmentation (16 Samples)

20025519

包含用于制备16个文库的试剂。不包含富集试剂。单独购买富集探针panel、Illumina纯化磁珠和标签接头。富集试剂只能与文库制备试剂(Illumina DNA Prep with Enrichment, (S) Tagmentation(16样本))一起订购。

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Panel (5)

TruSight Cancer – Enrichment Oligos only (4 or 8 Enrichment Reactions)

FC-121-0202

TruSight Cancer能靶向与多种癌症易感性相关的94个基因和284个SNP,与Nextera Flex for Enrichment联合使用时,为单一杂交的工作流程,足够进行8次富集反应,与TruSight Rapid Capture联合使用时足够进行4次富集反应。文库制备、富集和标签接头试剂需单独订购。

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TruSight Hereditary Cancer – Enrichment Oligos Only (8 Enrichment Reactions)

20029551

TruSight Hereditary Cancer——仅含富集寡核苷酸(8次富集反应)

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TruSight One – Enrichment Oligos only (6 Enrichment Reactions)

20029227

与Illumina DNA Prep with Enrichment, (S) Tagmentation搭配使用时,足以进行六次富集反应。单独购买文库制备试剂、富集试剂和标签接头。

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TruSight One Expanded – Enrichment Oligos only (6 Enrichment Reactions)

20029226

与Illumina DNA Prep with Enrichment, (S) Tagmentation搭配使用时,足以进行六次富集反应。单独购买文库制备和富集试剂以及标签接头。

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Illumina Custom Enrichment Panel (8 Enrichment Reactions)

20025371

与Illumina DNA Prep with Enrichment搭配使用时,足够进行8次富集反应。使用DesignStudio在2K–67K探针之间实现定制panel设计。单独购买文库制备和富集试剂以及标签接头。

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定制

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标签接头 (8)

Illumina® DNA/RNA UD Indexes Set A, Tagmentation (96 Indexes, 96 Samples)

20091654

Illumina® DNA/RNA UD Indexes Set A, Tagmentation(96个标签,96个样本)包括96个10 bp标签,足以标记96个样本。单独购买文库制备和富集试剂以及探针panel。

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Illumina® DNA/RNA UD Indexes Set B, Tagmentation (96 Indexes, 96 Samples)

20091656

Illumina® DNA/RNA UD Indexes Set B, Tagmentation(96个标签,96个样本)包括96个10 bp标签,足以标记96个样本。单独购买文库制备和富集试剂以及探针panel。

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Illumina® DNA/RNA UD Indexes Set C, Tagmentation (96 Indexes, 96 Samples)

20091658

Illumina® DNA/RNA UD Indexes Set C, Tagmentation(96个标签,96个样本)包括96个10 bp标签,足以标记96个样本。单独购买文库制备和富集试剂以及探针panel。

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Illumina® DNA/RNA UD Indexes Set D, Tagmentation (96 Indexes, 96 Samples)

20091660

Illumina® DNA/RNA UD Indexes Set D, Tagmentation(96个标签,96个样本)包括96个10 bp标签,足以标记96个样本。单独购买文库制备和富集试剂以及探针panel。

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IDT® for Illumina® DNA/RNA UD Indexes Set A, Tagmentation (96 Indexes, 96 Samples)

20027213

包含96、10 bp标签,足以标记96个样本。单独购买文库制备和富集试剂以及探针panel。

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IDT® for Illumina® DNA/RNA UD Indexes Set B, Tagmentation (96 Indexes, 96 Samples)

20027214

包含96、10 bp标签,足以标记96个样本。单独购买文库制备试剂、富集试剂和探针panel。

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IDT® for Illumina® DNA/RNA UD Indexes Set C, Tagmentation (96 Indexes, 96 Samples)

20042666

包含96、10 bp标签,足以标记96个样本。单独购买文库制备和富集试剂以及探针panel。

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IDT® for Illumina® DNA/RNA UD Indexes Set D, Tagmentation (96 Indexes, 96 Samples)

20042667

包含96、10 bp标签,足以标记96个样本。单独购买文库制备试剂、富集试剂和探针panel。

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服务 (1)

Illumina DNA Prep with Enrichment Training

20028457

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配套产品 (7)

Illumina Purification Bead, 100mL

20060057

包含一个100 mL微珠容器,用于文库纯化和大小选择。单独购买文库制备和富集试剂、探针panel和标签。

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Illumina Purification Bead, 400mL

20060058

包含四个100 mL微珠容器,用于文库纯化和大小选择。单独购买文库制备和富集试剂、探针panel和标签。

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Flex Lysis Reagent Kit (96 reactions)

20018706

包含使用Illumina DNA Prep和Illumina DNA Prep with Enrichment处理血液样本的试剂。单独购买文库制备、富集、富集探针panel和标签接头试剂。

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Infinium FFPE QC Kit (384 reactions)

WG-321-1001

Infinium FFPE QC Kit是使用Nextera Flex for Enrichment鉴定FFPE样本所需的配套产品。文库制备、富集、富集探针panel以及标签接头试剂需单独订购。

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Infinium MIDI Heatblock Insert

BD-60-601

Nextera Flex for Enrichment杂交步骤所需的SciGene Hybex微型加热系统的加热插块。

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Illumina® Free Adapter Blocking Reagent (12 Reactions)

20024144

该试剂在文库制备过程中使用,可降低标签跳跃水平。

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Illumina® Free Adapter Blocking Reagent (48 Reactions)

20024145

该试剂在文库制备过程中使用,可降低标签跳跃水平。

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选择汇总

产品

数量

Unit Price

常见问题解答

Illumina DNA Prep with Enrichment试剂盒兼容10–1000 ng或50–1000 ng FFPE的高质量双链基因组DNA(gDNA)起始量。对于人gDNA样本与其他复杂的大型基因组,建议的最小gDNA起始量为50 ng。

该试剂盒使用基于磁珠的转座酶复合物,用接头对基因组DNA进行片段化和标记,然后进行PCR步骤,将带标签的接头序列添加到DNA片段的末端。文库制备完成后,通过杂交生物素化的寡核苷酸探针并使用链霉亲和素磁珠(SMB)捕获靶向文库片段来富集靶标。从磁珠中洗脱捕获的标签文库,并在测序前扩增。

Illumina不支持或不支持将冷冻或凝固血液与Flex Lysis Reagent Kit一起使用,将血液裂解方案与Illumina DNA Prep with Enrichment一起使用。

Illumina DNA Prep with Enrichment的血液裂解实验方案针对K2-EDTA采集管(由Becton Dickinson生产)进行了优化。其他采血管可能有效,但尚未经过验证。

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订购NovaSeq X系列

将先进的化学、光学和信息学技术相结合,实现了超凡的速度和数据质量、出色的通量和可扩展性。

MiSeq Reagent Kit v3

与早期版本相比,优化的试剂盒可增加簇密度和读长,从而提高测序质量分值。

TruSight Oncology 500

使用涵盖所有主要变异类别以及来自FFPE组织的基因特征(TMB、MSI和HRD)的大型泛癌panel实现CGP。

Illumina DNA Prep

快速、集成的文库制备工作流程,适用于各种测序应用。

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