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Whole genome sequencing

Succeeding in your whole genome sequencing (WGS) research is critical to improving the lives of all living things—today and well into the future. With our xGen whole genome sequencing solutions, you can accelerate your research.

xGen™ NGS—made to reach.

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.

Icons Library_White Outline_85x85_Extraction

Extraction

Icons Library_White Outline_85x85_Library prep

LIBRARY PREP

xGen DNA Library Prep Kit EZ (16 or 96 rxn)
for high quality DNA samples

UDI primers (16 rxn)
For 8 or 10-base pair indexes

UDI Normalase primers (96 rxn)

Icons Library_White Outline_85x85_Normalase

Normalization

xGen Normalase Module (included in the 96 rxn size only)

Icons Library_White Outline_85x85_Sequence and analyze

Sequencing & analysis

IDT ALIGN Program

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.

Figure 1. Normalase enables streamlined library balancing and pooling process without the need to quantify samples and maximizes sequencing efficiency. xGen DNA library prep EZ libraries were generated with stubby adapters from 100 ng of NA12878 Coriell gDNA. Twelve library subsets were either pooled and normalized based on qPCR quantification or pooled and normalized using the xGen Normalase Module, followed by co-sequencing to determine percent reads identified of each index using (MiSeq® V2 Standard 300-cycle kit).

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*RUO—For research use only. Not for use in diagnostic procedures. Unless otherwise agreed to in writing, IDT does not intend for these products to be used in clinical applications and does not warrant their fitness or suitability for any clinical diagnostic use. Purchaser is solely responsible for all decisions regarding the use of these products and any associated regulatory or legal obligations.

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