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GeneRead Library Prep Kits

制备DNA文库用于NGS应用
  • 快速的单管式操作可节约多达50%的时间
  • 从少量起始材料中获得高产量
  • 可选的高保真扩增步骤可进行无偏差扩增
  • 高通量多重扩增
GeneRead Library Prep Kits可为NGS应用制备文库,工作流程经过优化,十分高效。该试剂盒能够快速、高效的制备DNA文库,制备的文库可用于Illumina或Life Technologies的Ion Torrent NGS平台。
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产品 货号 目录价:
QIAseq Ultralow Input Library Kit (12)
For 12 reactions: buffers and reagents for Ultralow Input End Polishing, Ultralow Input Ligation and HiFi library amplification. 12-plex adapters sold separately. For use with Illumina instruments.
180492
QIAseq Ultralow Input Library Kit (96)
For 96 reactions: buffers and reagents for Ultralow Input End Polishing, Ultralow Input Ligation and HiFi library amplification. Includes 96-plex Adapter Plate with individually pierceable foil sealed wells. For use with Illumina instruments.
180495
QIAseq FX DNA Library Kit (24)

Buffers and reagents for DNA fragmentation (including end repair and A-addition), ligation and library amplification; for use with Illumina instruments; includes a plate containing 24 adapters with different barcodes (pierceable foil seal, allowing usage of defined parts of plate)

180473
QIAseq FX DNA Library Kit (96)

Buffers and reagents for DNA fragmentation (including end repair and A-addition), ligation and library amplification; for use with Illumina instruments; includes a plate containing 96 adapters with different barcodes (pierceable foil seal, allowing usage of defined parts of plate)

180475
QIAseq 1-Step Amplicon Library Kit (12)

For 12 reactions: 1-Step Amplicon Enzyme Mix, 4x 1-Step Amplicon Buffer, Primer Mix Illumina Library Amp, HiFi PCR Master Mix, RNase-Free Water. Adapters sold separately.

180412
QIAseq 1-Step Amplicon Library Kit (96)

For 96 reactions: 1-Step Amplicon Enzyme Mix, 4x 1-Step Amplicon Buffer, Adapter Plate 96-plex Illumina, Primer Mix Illumina Library Amp, HiFi PCR Master Mix, RNase-Free Water

180415
GeneRead DNA Library I Kit (96)

Buffers and reagents for end-repair, A-addition, ligation and library amplification; for use with Illumina instruments; includes a plate containing 96 adapters with different barcodes (pierceable foil seal allowing usage of defined parts of plate)

180435
GeneRead DNA Library I Core Kit (12)
For 12 reactions: Buffers and reagents for end-repair, A-Addition, and ligation, for use with Illumina instruments
180432
GeneRead DNA Library I Core Kit (48)
For 48 reactions: Buffers and reagents for end-repair, A-Addition, and ligation, for use with Illumina instruments
180434
GeneRead DNA I Amp Kit (100)
For 100 reactions: Ready-to-use library amplification master mix and primer mix, for use with Illumina instruments
180455
GeneRead Adapter I Set 1-plex (12)
For 12 reactions: Adapters for DNA ligation, for use with Illumina instruments
180912
GeneRead Adapter I Set A 12-plex (144)
For 144 reactions: 12 barcoded adapters for ligation to DNA library, for use with Illumina instruments
180985
GeneRead Adapter I Set B 12-plex (144)
For 144 reactions: 12 barcoded adapters for ligation to DNA library, for use with Illumina instruments
180986
GeneRead DNA Library L Core Kit (12)
For 12 reactions: Buffers and reagents for end-repair, ligation and nick-repair, for use with Ion Torrent instruments
180462
GeneRead DNA L Amp Kit (100)
For 100 reactions: Ready-to-use library amplification master mix and primer mix, for use with Ion Torrent instruments
180485
GeneRead Adapter L Set 1-plex (12)
For 12 reactions: Adapters for DNA ligation, for use with Ion Torrent instruments
180922
GeneRead Adapter L Set 12-plex (72)
For 72 reactions: 12 barcoded adapters for ligation to DNA library, for use with Ion Torrent instruments
180994

GeneRead Library Prep Kits适用于分子生物学应用。该产品不适用于疾病的诊断、预防或治疗。


0
文库DNA的产量高。
使用Covaris仪器从E. Coli菌株DH10B中剪切基因组DNA(1 μg),使用GeneRead Library Prep (I) Kit或其他供应商的试剂盒制备Illumina兼容的DNA文库。使用18 μl Buffer EB洗脱文库,利用定量PCR测定文库的浓度。
1
高保真扩增,错配率低,序列偏差小。
将高度富含GC的Bordetella pertussis gDNA(GC含量67.7%)和高度富含AT的Streptobacillus moniliformis gDNA(GC含量26.3%)混合,使用GeneRead Library (I) Core Kit制备Illumina兼容的DNA文库,并使用GeneRead Library (I) Amp Kit(包含GeneRead HiFi Polymerase)或Supplier K的试剂盒进行扩增。使用Illumina MiSeq仪器测序扩增文库,并利用Galaxy平台分析保真度和序列覆盖度。GeneRead Library (I) Amp Kit的错误率更低,累积序列覆盖度更高,说明其文库扩增效果优于Supplier K的试剂盒。
2
准确的片段大小选择。
GeneRead Size Selection Kit可在文库制备后高效去除接头二聚体和接头单体。插图中显示了按比例缩放的数据图,图中显示了在进行大小选择后Illumina兼容的文库片段的正确大小分布。FU:荧光单位。
3
覆盖更完整的序列。
将高度富含GC的Bordetella pertussis gDNA(GC含量67.7%)和高度富含AT的Streptobacillus moniliformis gDNA(GC含量26.3%)混合,使用GeneRead Library Prep (I) Kit制备Illumina兼容的DNA文库,并使用GeneRead Library Amp (I) Kit(包含GeneRead HiFi Polymerase)或Supplier K的试剂盒进行扩增。使用Illumina MiSeq仪器测序扩增文库,并利用Galaxy平台分析保真度和序列覆盖度。GeneRead Library Prep Kit在富含GC和AT的DNA区域具有较Supplier K的试剂盒更高的序列覆盖度。
4
文库DNA产量高,覆盖分布均匀。
[A] 使用Covaris仪器剪切基因组DNA(50 ng),并使用GeneRead Library Prep (I) Kit制备Illumina兼容的DNA文库。使用Illumina MiSeq仪器测序的结果显示,中间覆盖度为49倍,覆盖度分布均匀。[B] E. coli菌株DH10B中剪切基因组DNA(1 μg),并使用GeneRead Library Prep (L) Kit或其他供应商的试剂盒生成Ion Torrent兼容的DNA文库。扩增10个循环后,在Ion Torrent PGM仪器上对两种文库进行测序,分析累积的标准化覆盖度。
5
优化的工作流程,制备与Illumina兼容的DNA文库。
Library Prep (I) Kit采用优化的单管实验方案,回收步骤较少,包括可选的高保真文库扩增。上述文库DNA制备所需的操作时间和总时间较Supplier I的文库制备系统明显缩短。
6
优化的工作流程,制备与Ion Torrent兼容的DNA文库。
GeneRead Library Prep (L) Kit采用优化的单管式步骤,回收步骤较少,包括可选的高保真文库扩增。上述文库DNA制备所需的操作时间和总时间较Supplier L的文库制备系统明显缩短。
Performance
文库DNA产量高,位点覆盖全面
GeneRead Library Prep Kits的工作流程经过优化,十分高效,能够稳定获得高产量DNA文库,序列偏差小,错配率低;文库适合用于Illumina或Life Technologies的Ion Torrent NGS平台。高效的连接反应和可选的高保真扩增步骤能够确保从少量起始材料、甚至是50 ng起始材料中获得高产量文库,且位点覆盖全面(参见High yields of library DNA with uniform coverage distributionSuperior yields of library DNA)。

错配率低,序列偏差小
为确保从少量起始材料中获得最大产量,GeneRead Library Prep Kits含有创新的高保真预混液,其中含有用于扩增的GeneRead HiFi Polymerase。GeneRead HiFi Polymerase是独特的、高度特异性的进行性聚合酶,能够准确扩增文库DNA,错配率低,序列偏差小(参见Low error rates with minimal sequence bias due to high-fidelity amplification)。

扩增富含GC和AT的序列时,序列偏差较小
在标准PCR扩增中,AT或GC含量高的DNA序列扩增产物可能较少或没有,导致错误的序列数据或NGS测序结果。GeneRead HiFi Polymerase与预混液中独特的配方共同作用,能够确保均一扩增GC含量高的基因组序列,确保在后续的测序实验中获得准确结果(参见Better sequence coverage)。
Principle
GeneRead Library Prep Kits的单管式工作流程经过优化,能够稳定获得高产量DNA文库;文库适合用于Illumina或Life Technologies的Ion Torrent NGS平台。与其他供应商的文库制备工作流程相比,快速的单管式实验流程采取较少的回收步骤,且包含可选的高保真文库扩增步骤。GeneRead Library Prep Kits极大程度减少了手工操作可能带来的污染和失误风险,节省时间和人力(参见Optimized workflow for Illumina-compatible DNA librariesOptimized workflow for Ion Torrent-compatible DNA libraries)。
Procedure
制备适用于Illumina平台的DNA文库的实验流程
由长片段DNA组成的样本首先被剪切为300 bp(使用Illumina MiSeq仪器,version 1)或500 bp(使用Illumina MiSeq仪器,version 2)的中等长度片段的随机文库。剪切后,修复DNA片段两端,在每条链的3'端加上Poly A尾巴。之后,将扩增和测序所必须的接头连接到DNA片段两端。GeneRead Library Prep (I) Kit中含有的Barcode接头含有独特的辨别序列,因此能够进行多重测序反应。之后选择一定长度的片段进行纯化。可使用GeneRead Size Selection Kit(货号180514)进行准确的片段回收和片段长度选择(参见Precise size selection)。为确保从少量起始材料中获得高产量,可选择性进行高保真扩增步骤。

制备适用于Ion Torrent平台的DNA文库的实验流程
由长片段DNA组成的样本首先被剪切为400 bp(使用Ion Torrent PGM仪器)或200 bp(使用Ion Proton仪器)的中等长度片段的随机文库。剪切后,修复DNA片段两端。之后,将扩增和测序所必须的接头连接到DNA片段两端。GeneRead Library Prep (L) Kit中含有的Barcode接头含有独特的辨别序列,因此能够进行多重测序反应。之后选择一定长度的片段进行纯化。为确保从少量起始材料中获得高产量,可选择性进行高保真扩增步骤。
Applications
GeneRead Library Prep Kits能够快速、高效的制备DNA文库,制备的文库可用于Illumina或Life Technologies的Ion Torrent NGS平台。

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试剂盒操作手册 (5)

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For preparation of DNA libraries for next-generation sequencing (NGS) applications that use Illumina instruments
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For preparation of DNA libraries for next-generation sequencing (NGS) applications that use Ion Torrent instruments from Life Technologies
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其他资源 (1)
For the preparation, size selection, and purification of DNA libraries for NGS applications
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