Whole genome sequencing

Overview

• Recommended products—curated for whole genome sequencing
• Short workflow—create a normalized library pool in approximately 4 hours
• Low input, high complexity—up to 3x’s fewer duplicates from 1 ng of DNA
• Enzymatic fragmentation—no need for specialized equipment for library preparation
• xGen Normalase™ technology—a proprietary enzymatic method of library normalization for multiplexing applications
• Automation friendly—for a variety of platforms

Whole genome sequencing provides a comprehensive set of genomic sequence data about an organism, tissue, or even metagenomic DNA samples. In comparison to targeted sequencing, WGS decodes each of the fragments in the NGS indexed libraries. The sequence data is then aligned to a reference genome or assembled into contigs for de novo genome assemblies.

For low-throughput labs, the 16-reaction product solution for whole genome sequencing includes library prep and adapters. For library prep, the xGen DNA Library Prep Kit EZ uses an enzymatic fragmentation strategy to shear high-quality DNA samples. The kit employs ligation-based library prep, and it includes enzymes for fragmentation, end-repair, a high-fidelity polymerase for indexing PCR, and A‑tailing, as well as a ligase for the addition of the xGen Stubby Adapters (included). The product solution offers premixed xGen UDI Primer Pairs for indexing by PCR, which generates a NGS library ready for Illumina® sequencing.

For high-throughput labs, the 96-reaction size includes the xGen Normalase Module, a proprietary enzymatic library normalization kit for multiplexing. The method eliminates the need for individual sample quantification followed by manual equimolar pooling. Instead, the fragments are indexed by PCR using the included plate of xGen Normalase UDI Primer Pairs followed by the xGen Normalase Module. Final normalized library pools are then ready for Illumina^®-based multiplex sequencing.

Method data

Normalase allows for the formation of balanced library pools without quantification resulting in uniform sequencing data

Normalization using the proprietary Normalase workflow in comparison to the traditional qPCR quantification method provides more uniform results, maximizing the value of the sequencing data as it provides a much tighter grouping of reads between samples.

In an experiment where the xGen DNA Library Prep Kit EZ was used in conjunction with NA12878 gDNA, stubby adapter, xGen UDI primers, and Normalase, the coefficient of variation (CV) was lower (6.9%) compared than when libraries were quantified by qPCR (18%, Figure 1). Meaning that the use of Normalase allows for a faster, more efficient workflow for multiplex sequencing.

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