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Whole Genome Sequencing

As next-generation sequencing (NGS) costs decrease and bioinformatics analysis pipelines improve, the utility of whole genome sequencing (WGS) is ever expanding. However, as WGS sample throughput increases, preparing high quality sequenceable libraries from a wide range of sample types in a timely manner is a bottleneck impeding sequencing productivity.

Whether you are new to NGS or your capabilities already include high throughput automated WGS, our Accel-NGS® 2S DNA Library Kits can help you maximize your data output, giving you the best genome-wide coverage of any kit on the market, and reducing your sequencing costs.

SWIFT PRODUCT LINES COMPATIBLE WITH WHOLE GENOME SEQUENCING:

Accel-NGS 2S Plus DNA Library Kit

The Swift advantage:
  • Proprietary adapter ligation minimizes bias and supports inputs as low as 10 pg.
  • Superior library complexity generates more unique library molecules.
  • Minimal base composition bias from AT-/GC-rich sequences allows analysis of challenging genomes.

Accel-NGS 2S PCR-Free DNA Library Kit

The Swift advantage:
  • PCR-free library preparation from inputs as low as 10 ng.
  • Superior library complexity generates more unique library molecules.
  • Minimal base composition bias from AT-/GC-rich sequences allows analysis of challenging genomes.

A Superior Library Kit

The Accel-NGS 2S DNA Library Kits utilize a proprietary adapter attachment enzymology that first maximizes the number of available ends for ligation with two dedicated repair steps that repair both ends of both strands of each DNA fragment. Then, two ligation steps sequentially add adapter sequences to the 5’ and 3’ repaired ends. This approach leads to highly efficient conversion of DNA fragments into library molecules, allowing PCR-free libraries to be generated from 100 ng of starting material, or as little as 10 ng of starting material when pooling samples or using circulating, cell-free DNA from plasma. In addition, the template-independent adapter attachment chemistry results in complex libraries that faithfully represent the base composition of starting material.

Increased Conversion of Input DNA Reduces Sequencing Costs

The chemistry powering the Accel-NGS 2S DNA Library Kits enables capture of a more diverse set of molecules than any other leading kit. The high complexity set of library molecules leads to more uniquely mapped reads, better representation of diverse regions, and consequently more uniform sequence coverage. In turn, more uniform coverage reduces or eliminates missing sections of the genome from the sequencing data set while simultaneously leading to lower sequencing costs.

WGS-complexity

Library complexity was obtained at various sequencing depths for Accel-NGS 2S libraries compared to libraries made with the leading kit.

Libraries That Reflect Your NGS Samples

Extreme AT/GC base composition sequences adversely affect adapter ligation efficiency for most library methods, reducing complexity of the library and evenness of coverage obtained by whole genome sequencing. Accel-NGS 2S PCR-Free WGS libraries demonstrate complete coverage across the genome close to the theoretical average coverage predicted by Poisson statistical distribution and relative coverage close to 1.0 at nearly all base compositions. This means that regardless of the AT/GC base composition, the Accel-NGS 2S Kit chemistry can convert the template DNA to library molecules, improving coverage.

To demonstrate this, the Accel-NGS 2S PCR-Free DNA Library Kit was tested with the B. pertussis genome (68% GC) and the P. falciparum genome (19% GC). These results reveal balanced, near-theoretical coverage of these extreme microbial genomes. The same can be seen in regions of the human genome where base composition is skewed toward AT-/GC-rich sequences. The Accel-NGS 2S Kit chemistry effectively captures and converts these template molecules with the same conversion efficiency as less difficult regions.

wgs-extreme-covg_rev3

PCR-free libraries were made from 500 ng samples. Shown are the GC content of each read vs. theoretical in silico distribution (a = B. Pertussis, c = P. falciparum 3D7) and the normalized coverage vs. GC content (b = B. Pertussis, d = P. falciparum 3D7).

Convert and Sequence Difficult FFPE Samples

WGS-ffpe-covg

Translational research FFPE libraries from 10 ng (left) or 1 ng (right) followed by 9 & 12 cycles of PCR. Sequencing was performed on Illumina HiSeq 2500. Picard CollectGcBiasMetrics demonstrated evenness of coverage regardless of input amount and number of PCR cycles.

Human DNA from FFPE samples is susceptible to substantial damage resulting from fixation processes that introduce cross-links and fragmentation; oxidation damage also accumulates during prolonged storage. This damage can greatly reduce the number of usable DNA molecules for NGS library preparation, becoming a barrier to analysis. Accel-NGS 2S Plus and Accel-NGS 2S PCR-Free technology addresses the growing demand for a method that can convert a small amount of FFPE DNA into a high quality WGS library.

Unlike many competing products, which are not recommended for use with FFPE, Accel-NGS 2S chemistry can handle both damaged and short DNA fragments. This makes Accel-NGS 2S ideally suited for translational samples such as FFPE or plasma.