QIAseq FX DNA Library Kit

QIAseq FX DNA library preparation kits are designed for fast, enzymatic preparation of high-complexity whole-genome libraries with low GC bias. Fragment size, input amount (20 pg –1 µg) and batch size are customizable, making the kit highly suited for next-generation sequencing (NGS) projects. The all-enzymatic DNA fragmentation takes just 2.5 hours and requires no DNA shearing equipment, saving substantial time and costs.

Without the use of expensive DNA shearing equipment, substantial time and costs are saved. High-fidelity amplification can be added to the otherwise PCR-free protocol to increase library yield. The streamlined, optimized protocol can be automated, reducing hands-on time and run-to-run variability. The technology is compatible with Illumina sequencing instruments, providing configurations for up to 1536 samples. 

The NGS library preparation kit supports researchers working with limited or degraded DNA – including low-input clinical, environmental and microbial samples – where mechanical shearing is impractical or causes sample loss. The kit is equally suited for core labs requiring consistent, automated library preparation across human whole genome sequencing (WGS), metagenomic library preparation and microbial genome sequencing projects. 

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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.

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.

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.

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. 

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