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。
Product Cat. no. List price:
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
£245.00
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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
£2,251.00
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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 combinatorial dual index adapters with different bar codes (pierceable foil seal, allowing usage of defined parts of plate)

180473
£658.00
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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 combinatorial dual index adapters with different bar codes (pierceable foil seal, allowing usage of defined parts of plate)

180475
£2,605.00
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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
£237.00
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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
£2,135.00
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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
£799.00
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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
£268.00
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GeneRead Adapter I Set 12-Plex (72)
For 72 reactions: 12 barcoded adapters for ligation to DNA library, for use with Illumina instruments
180984
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GeneRead Library Prep Kits are intended for molecular biology applications. These products are not intended for the diagnosis, prevention, or treatment of a disease.
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Superior yields of library DNA.|Low error rates with minimal sequence bias due to high-fidelity amplification.|Precise size selection.|Better sequence coverage.|High yields of library DNA with uniform coverage distribution.|Optimized workflow for preparation of Illumina-compatible DNA libraries.|Optimized workflow for generation of Ion Torrent-compatible DNA libraries.|
Genomic DNA (1 μg) from E. Coli strain DH10B was sheared using a Covaris instrument and made into an Illumina–compatible DNA library using the GeneRead Library Prep (I) Kit or a kit from another supplier. The library was eluted in 18 μl Buffer EB and library concentrations were determined by quantitative PCR.|A mixture of highly GC-rich Bordetella pertussis gDNA (GC content 67.7%) and highly AT-rich Streptobacillus moniliformis gDNA (GC content 26.3%) was pooled, made into an Illumina-compatible DNA library using the GeneRead Library (I) Core Kit, and amplified with the GeneRead Library (I) Amp Kit, which contains GeneRead HiFi Polymerase, or a kit from Supplier K. The amplified libraries were sequenced using the Illumina MiSeq instrument, and fidelity and sequence coverage were analyzed using the Galaxy platform. Low error rates and greater cumulated sequence coverage demonstrate that the GeneRead Library (I) Amp Kit provides superior library amplification compared to the kit from Supplier K.|The GeneRead Size Selection Kit effectively removes adapter dimers and adapter monomers following library preparation. A scaled-up image of the above data showing the correct size distribution of Illumina-compatible library fragments following size selection is shown in inset. FU: Fluorescence units.|A mixture of highly GC-rich Bordetella pertussis gDNA (GC content 67.7%) and highly AT-rich Streptobacillus moniliformis gDNA (GC content 26.3%) was pooled, made into an Illumina-compatible DNA library using the GeneRead Library Prep (I) Kit, and amplified with the GeneRead Library Amp (I) Kit, which contains GeneRead HiFi Polymerase, or a kit from Supplier K. The amplified libraries were sequenced using the Illumina MiSeq instrument, and fidelity and sequence coverage was analyzed using the Galaxy platform. The GeneRead Library Prep Kit provided greater sequence coverage in GC- and AT-rich areas of DNA, compared to the kit from Supplier K.|[A] Genomic DNA (50 ng) was sheared using a Covaris instrument and made into an Illumina-compatible DNA library using the GeneRead Library Prep (I) Kit. Sequencing using an Illumina MiSeq instrument revealed a median coverage of 49-fold with uniform coverage distribution. [B] Genomic DNA (1 μg) from E. coli strain DH10B was sheared and used to generate an Ion Torrent-compatible DNA library using the GeneRead Library Prep (L) Kit or a kit from another supplier. After amplification for 10 cycles, both libraries were sequenced on an Ion Torrent PGM instrument and the cumulated normalized coverage was analyzed.|Library Prep (I) Kit uses an optimized one-tube protocol, with fewer cleanup steps and optional high-fidelity library amplification. The hands-on time and total time required for preparation of library DNA is significantly reduced compared to the library preparation system from Supplier I.|The GeneRead Library Prep (L) Kit uses an optimized, one-tube procedure, with fewer cleanup steps and optional high-fidelity library amplification. The hands-on time and total time required for preparation of library DNA is significantly reduced compared to the library preparation system from Supplier L.|
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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Additional Resources
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For the preparation, size selection, and purification of DNA libraries for NGS applications
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The GeneRead Library Prep Kit delivers high yields of library DNA.
Superior yields of library DNA.
Genomic DNA (1 μg) from E. Coli strain DH10B was sheared using a Covaris instrument and made into an Illumina–compatible DNA library using the GeneRead Library Prep (I) Kit or a kit from another supplier. The library was eluted in 18 μl Buffer EB and library concentrations were determined by quantitative PCR.
Low error rates with minimal sequence bias due to high-fidelity amplification.
Low error rates with minimal sequence bias due to high-fidelity amplification.
A mixture of highly GC-rich Bordetella pertussis gDNA (GC content 67.7%) and highly AT-rich Streptobacillus moniliformis gDNA (GC content 26.3%) was pooled, made into an Illumina-compatible DNA library using the GeneRead Library (I) Core Kit, and amplified with the GeneRead Library (I) Amp Kit, which contains GeneRead HiFi Polymerase, or a kit from Supplier K. The amplified libraries were sequenced using the Illumina MiSeq instrument, and fidelity and sequence coverage were analyzed using the Galaxy platform. Low error rates and greater cumulated sequence coverage demonstrate that the GeneRead Library (I) Amp Kit provides superior library amplification compared to the kit from Supplier K.
Precise size selection
Precise size selection.
The GeneRead Size Selection Kit effectively removes adapter dimers and adapter monomers following library preparation. A scaled-up image of the above data showing the correct size distribution of Illumina-compatible library fragments following size selection is shown in inset. FU: Fluorescence units.
Better sequence coverage and minimal bias when amplifying GC- or AT-rich sequences
Better sequence coverage.
A mixture of highly GC-rich Bordetella pertussis gDNA (GC content 67.7%) and highly AT-rich Streptobacillus moniliformis gDNA (GC content 26.3%) was pooled, made into an Illumina-compatible DNA library using the GeneRead Library Prep (I) Kit, and amplified with the GeneRead Library Amp (I) Kit, which contains GeneRead HiFi Polymerase, or a kit from Supplier K. The amplified libraries were sequenced using the Illumina MiSeq instrument, and fidelity and sequence coverage was analyzed using the Galaxy platform. The GeneRead Library Prep Kit provided greater sequence coverage in GC- and AT-rich areas of DNA, compared to the kit from Supplier K.
High yields of library DNA with uniform coverage distribution.
High yields of library DNA with uniform coverage distribution.
[A] Genomic DNA (50 ng) was sheared using a Covaris instrument and made into an Illumina-compatible DNA library using the GeneRead Library Prep (I) Kit. Sequencing using an Illumina MiSeq instrument revealed a median coverage of 49-fold with uniform coverage distribution. [B] Genomic DNA (1 μg) from E. coli strain DH10B was sheared and used to generate an Ion Torrent-compatible DNA library using the GeneRead Library Prep (L) Kit or a kit from another supplier. After amplification for 10 cycles, both libraries were sequenced on an Ion Torrent PGM instrument and the cumulated normalized coverage was analyzed.
Optimized workflow allows more rapid and efficient preparation of Illumina-compatible DNA libraries.
Optimized workflow for preparation of Illumina-compatible DNA libraries.
Library Prep (I) Kit uses an optimized one-tube protocol, with fewer cleanup steps and optional high-fidelity library amplification. The hands-on time and total time required for preparation of library DNA is significantly reduced compared to the library preparation system from Supplier I.
Optimized workflow allows more rapid and efficient preparation of Life Technologies-compatible DNA libraries.
Optimized workflow for generation of Ion Torrent-compatible DNA libraries.
The GeneRead Library Prep (L) Kit uses an optimized, one-tube procedure, with fewer cleanup steps and optional high-fidelity library amplification. The hands-on time and total time required for preparation of library DNA is significantly reduced compared to the library preparation system from Supplier L.