このソリューションをお試しになりませんか？
今すぐ当社のチームにご連絡の上、QIAseq FX DNA Library UDI Kit (24) トライアルキットの見積りをご依頼ください。
QIAseq FX DNA Library UDI Kit (24)
カタログ番号 / ID. 180477
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 unique dual index Y-adapters.
産物タイプ
Library Kit
Beads
Normalizer
インデックス数
24
96
384
タイプ
UDI
CDI
QIAseq FX DNA Library UDI Kit (24)は分子生物学的アプリケーション用であり、疾病の診断、予防、あるいは治療に使用することはできません。
このソリューションをお試しになりませんか？
今すぐ当社のチームにご連絡の上、QIAseq FX DNA Library UDI Kit (24) トライアルキットの見積りをご依頼ください。
特徴
- わずか2.5 時間の簡便なワークフロー
- バイアスを最小限にすることで、均一なカバレッジを取得可能
- Unique Dual Index（UDI）アダプター対応で、最大384 サンプルのマルチプレックスが可能
- 実験デザインに合わせて断片化サイズ、インプット量、サンプル処理数を柔軟に調整可能
- 20 pg～1 µg までの幅広いインプットDNA 量に対応
製品詳細
QIAseq FX DNA Library Kit は、酵素法に基づいた簡便なライブラリー調製試薬です。3ステップのシンプルなプロトコールにより、ラン間の誤差を最小限にします。本キットは、24または96サンプルフォーマットに対応しており、QIAseq Unique Dual Index Y-Adapter Kits と組合せることで最大384 サンプルのマルチプレックスを可能にします。（イルミナシークエンサー対応）
高額なDNA断片化装置は不要でコストパフォーマンスが高いライブラリー調製試薬をぜひお試しください。
パフォーマンス
Fast and efficient enzymatic library conversion
The fast and efficient workflow (see figure: Workflow) for enzymatic library conversion provided by the QIAseq FX DNA Library Kit is specifically designed for success in whole-genome sequencing for a wide range of applications.
| Feature | Specifics | Further details |
|---|---|---|
| Source genomic DNA from any organism | Oil, gut microbiome, wastewater and clinical isolates, etc. | Suitable for metagenomics and profiling whole-genome projects |
| Flexible input range | 20 pg – 1 μg genomic DNA | For low-DNA samples or for libraries that will be used in several downstream applications |
| Enzymatic fragmentation | Comparable to mechanical shearing | Fast 2.5 hour workflow Easily automated |
| Customizable fragment size | Highly reproducible fragmentation dependent on incubation time (see figures: Customizable fragment sizes and High reproducibility) |
|
| Similar total coverage depth for genomic targets | No additional sequencing for low-coverage targets | |
| Minimal GC bias | Improves representation in high- and low-GC microbial genomes (see figures: Minimal GC bias compared to other enzymatic methods) and No significant differences between coverage of high or low GC genomic regions) | |
| Coverage | High library complexity with low duplicate rates | Reduces the need for additional sequencing to bring low-coverage targets up to an interpretable coverage range, saving time and resources Maximizes interpretable data and overcomes common shortcoming of other enzymatic fragmentation methods (see figures: Superior coverage distribution and Lower duplication rate) |
| PCR-free workflow | Adapter ligation chemistry and dual bar coded adapters when PCR amplification is not needed | Provides superior data quality for NGS sequencing of whole-genome libraries |
原理
The novel nuclease formulation, all-enzymatic fragmentation and protocol are optimized for microbial genomes and shotgun (e.g., whole genome, metagenomic) sequencing workflows on Illumina platforms. The fragment size is adjustable simply by changing incubation time of the dsDNA with the nuclease. Optimized enzyme and buffer compositions ensure high sequencing library yield. Streamlined library construction protocols also enable straightforward automation. The fast, fully enzymatic procedure – from DNA fragmentation to NGS library – requires no cleanup steps until after adapters have been ligated to the sample DNA.
操作手順
Overview
The procedure detailed below consists of three, easy-to-follow steps starting from genomic DNA and ending with sequencer-ready, whole-genome libraries. Dual bar-coded, plate-format adapters are included with the kit. Each well contains a unique combination of two identification bar codes. Up to 1536-plex pooling is possible prior to sequencing.
| Input | Step | Output |
|---|---|---|
| Purified genomic DNA | FX enzymatic fragmentation of genomic or long-range DNA. Fragment size is controlled by the time of the reaction. Sequence-agnostic fragmentation. | DNA fragments of specified size |
| Fragmented DNA | In an end-repair reaction, an A is added to the 3′ ends of the fragmented DNA. | Fragments ready for adapter ligation |
| Platform-specific dual bar coded adaptors are ligated to both end of the DNA fragments; no cleanup necessary. The PCR-free workflow ends here. | Sequencer-ready dual bar-coded whole-genome libraries | |
| Amplification (optional) | In the optional amplification step (only necessary for the PCR workflow), DNA regions with vastly different GC contents are evenly amplified, minimizing sequencing bias caused by PCR. | Increased library yield |
Cleanup and removal of adapter-dimers
Following library construction, the reaction cleanup and removal of adapter dimers can be achieved by using QIAseq Beads, enabling easy automation on various high-throughput automation platforms.
Library normalization
The QIAseq Library Normalizer seamlessly integrates with our protocol, removing the need for tedious qPCR and manual dilution of libraries before pooling. Normalized libraries are ready-to-sequence dsDNA at approximately 4 nmol/L.
アプリケーション
QIAseq FX DNA Library Kit generates high-quality whole-genome libraries appropriate for the following types of sequencing:
- Shotgun, such as whole genome (WGS), including PCR-free WGS
- Whole exome (WES), or other downstream hybrid capture
- Amplicon
- Metagenomic
These types of sequencing are used in the following applications:
- Environmental DNA analysis
- Microbiome profiling
- Clinical genomics research
- Microbial genome sequencing
- Population genetics studies
- Taxonomic profiling
- Low-biomass microbial studies
裏付けデータと数値
Workflow
Streamlined three-step workflow. Go from purified gDNA to sequencer-ready libraries in just 2.5 hours with the QIAseq FX DNA Library Kit.
リソース
Template Files (4)
Kit Handbooks (2)
Protocols (6)
Safety Data Sheets (1)
Certificates of Analysis (1)
Brochures and Guides (1)
出版物
Validation and standardization of DNA extraction and library construction methods for metagenomics-based human fecal microbiome measurements.
Microbiome; 2021; 9 (1):95 2021 Apr 29 PMID:33910647