DNA library preparation for ultra-low sample input: NxSeq

The NxSeq® UltraLow DNA library kits from LGC Biosearch/Lucigen enable the preparation of complex, unbiased DNA fragment libraries from extremely low DNA input amounts. These kits utilise high efficiency adaptor ligation to yield improved sequencing depth uniformity and better coverage with few missed regions, allowing the preparation of DNA fragment libraries from as little as 50 pg to as much as 100 ng of sheared or fragmented DNA. Featuring a quick and easy protocol, library preparation can be completed in just three hours, with each step optimised to ensure peak performance on Illumina® sequencers.

Benefits of using the NxSeq UltraLow DNA library kits
• High quality libraries from even ultra-low DNA inputs
• High efficiency adaptor ligation ensures fewer missed regions
• Quick and easy protocol - library prep in just 3 hours
• Compatible with limited samples including genomic DNA, FFPE, cell-free DNA or samples for exome-seq or ChIP-seq
• Optimised for Illumina® sequencers
• Complex, unbiased libraries created
• 12 or 96 reaction kits available, depending on throughput requirements
• Includes guidance for mechanical shearing and enzymatic fragmentation

Consistent, high quality sequencing data from 50 pg to 100 ng of genomic DNA

Sequencing data from DNA fragment libraries generated from 50 pg to 100 ng of sheared E. coli gDNA. Mechanically sheared (Covaris LE220, peak size 300 bp) E. coli genomic DNA was serially diluted and used to make triplicate libraries starting with the indicated gDNA input amounts and number of PCR amplification cycles using the NxSeq UltraLow DNA Library Kit v2 and a NxSeq Single Indexing Kit. The final libraries were quantitated and diluted to 2 nM based on Bioanalyzer® (size) and fluorescence-based DNA quantitation. The diluted libraries were then pooled and sequenced on a MiSeq® using 2 x 150 bp chemistry, and the average data from the triplicate libraries are shown.

NxSeq UltraLow DNA library kits feature high efficiency adaptor ligation

Two independent sets of libraries were prepped per kit/organism using manufacturer’s recommended protocols and 1 ng of the same sheared genomic DNA input per library. DNA fragments with adaptors ligated to both ends were measured using triplicate qPCR assays and an universal qPCR primer set, designed by Biosearch Technologies, that binds to and amplifies all adaptor-ligated DNA fragments independent of the kit used. Efficiency was determined by comparing the qPCR quantitation to fluorescence DNA quantitation. Efficiency data was averaged and then normalised to the corresponding NxSeq UltraLow Library Kit data (1.0) and plotted.

Applications
• Whole genome sequencing (WGS)
• Whole genome resequencing (WGR) for genomic assembly
• SNV/mutation or copy number variation (CNV) identification
• ChIP-seq
• Exome-seq

Material available for download
NxSeq UltraLow DNA Library Kit v2 (12 reactions) manual
NxSeq UltraLow DNA Library Kit v2 (96 reactions) manual

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