QIAseq and GeneRead Library Prep Kits

For fast and efficient preparation of DNA libraries for use in NGS applications
  • Fast, one-tube procedure that allows up to 50% time savings
  • High yields from minimal amounts of starting material
  • Unbiased amplification with an optional, high-fidelity amplification step
  • Multiplexing, high-throughput capability
GeneRead Library Prep Kits provide an optimized and efficient workflow for library preparation in next-generation sequencing (NGS) applications. The kits allow fast and efficient preparation of DNA libraries that are for use on NGS platforms from Illumina and the Ion Torrent instrument from Life Technologies. Procedures can be automated on the QIAcube.

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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
询价
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
询价
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
询价
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
询价
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
询价
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
询价
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
询价
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
询价
GeneRead Adapter I Set 12-Plex (72)
For 72 reactions: 12 barcoded adapters for ligation to DNA library, for use with Illumina instruments
180984
询价

QIAseq and 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。


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文库DNA的产量高。|高保真扩增,错配率低,序列偏差小。|准确的片段大小选择。|覆盖更完整的序列。|文库DNA产量高,覆盖分布均匀。|优化的工作流程,制备与Illumina兼容的DNA文库。|优化的工作流程,制备与Ion Torrent兼容的DNA文库。|
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。|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。|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。|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。|[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。|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。|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。|
性能
High yields of library DNA with uniform coverage distribution
GeneRead Library Prep Kits provide an efficient and optimized workflow to reproducibly generate high yields of DNA library, with minimal sequence bias and low error rates, for use on NGS platforms from Illumina or the Ion Torrent instrument from Life Technologies. Highly efficient ligation reactions and an optional, high-fidelity amplification step ensure superior library yields with uniform coverage distribution, even from as low as 50 ng starting material (see figures High yields of library DNA with uniform coverage distribution and Superior yields of library DNA).

Low error rates with minimal sequence bias
To ensure maximum yields from minimum amounts of starting material, GeneRead Library Prep Kits include an innovative high-fidelity master mix that includes GeneRead HiFi Polymerase for an optional amplification step. GeneRead HiFi Polymerase is a unique, highly specific and processive enzyme that delivers accurate amplification of library DNA with low error rates and minimum bias (see figure Low error rates with minimal sequence bias due to high-fidelity amplification).

Lower sequence bias when amplifying GC- and AT-rich regions
In standard PCR amplification procedures, regions of DNA with high AT or GC content can result in little or no amplification, leading to misleading sequence data and NGS results. The GeneRead HiFi Polymerase, together with its unique buffer formulation, ensures uniform amplification of genomic regions that contain highly variable GC content, thereby ensuring even coverage in subsequent sequencing reactions (see figure Better sequence coverage).
原理
GeneRead Library Prep Kits use an optimized, one-tube workflow to reproducibly generate high yields of DNA library for use on NGS platforms from Illumina or the Ion Torrent instrument from Life Technologies. The fast, one-tube procedure uses fewer cleanup steps than library preparation workflows from other suppliers and includes an optional, high-fidelity library amplification step. GeneRead Library Prep Kits save time and effort while minimizing the variability caused by handing, as well as the risk of contamination (see figures Optimized workflow for Illumina-compatible DNA libraries and Optimized workflow for Ion Torrent-compatible DNA libraries).
操作流程
Procedure for generating Illumina-compatible DNA libraries
Samples consisting of longer DNA fragments are first sheared into a random library of fragments that are a median fragment size of 300 bp (when using the Illumina MiSeq instrument, version 1), or 500 bp (when using the Illumina MiSeq instrument, version 2), in length. Following fragmentation, the ends of the DNA fragments are repaired and an A-overhang is added at the 3'-end of each strand. Afterwards, adaptors, which are necessary for amplification and sequencing, are ligated to both ends of the DNA fragments. Barcode adapters, which contain a unique identifying sequence, are also available with the GeneRead Library Prep (I) Kit and enable multiplex sequencing reactions to be performed. Fragments are then size selected and purified. Cleanup and size selection can be done using the GeneRead Size Selection Kit (cat. no. 180514), which allows precise, column-based size selection of DNA (see figure Precise size selection). To ensure maximum yields from minimum amounts of starting material, an optional high-fidelity amplification step can also be performed.

Procedure for generating Ion Torrent-compatible DNA libraries
Samples consisting of longer DNA fragments are first sheared into a random library of fragments that are a median fragment size of 400 bp (when using the Ion Torrent PGM instrument), or 200 bp (when using the Ion Proton instrument), in length. Following fragmentation, the ends of the DNA fragments are repaired. Afterwards, adaptors, which are necessary for amplification and sequencing, are ligated to both ends of the DNA fragments. Barcode adapters, which contain a unique identifying sequence, are also available with the GeneRead Library Prep (L) Kit and enable multiplex sequencing reactions to be performed. Fragments are then size selected and purified. To ensure maximum yields from minimum amounts of starting material, an optional high-fidelity amplification step can be performed.

应用
GeneRead Library Prep Kits allow fast and efficient preparation of DNA libraries that are optimized for use on NGS platforms from Illumina and the Ion Torrent instrument from Life Technologies.

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For the preparation, size selection, and purification of DNA libraries for NGS applications
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图片
The GeneRead Library Prep Kit delivers high yields of library DNA.
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。
Low error rates with minimal sequence bias due to high-fidelity amplification.
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。
Precise size selection
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。
Better sequence coverage and minimal bias when amplifying GC- or AT-rich sequences
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。
High yields of library DNA with uniform coverage distribution.
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。
Optimized workflow allows more rapid and efficient preparation of Illumina-compatible DNA libraries.
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。
Optimized workflow allows more rapid and efficient preparation of Life Technologies-compatible DNA libraries.
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。