Twist Human Methylome Panel | Twist Bioscience

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The Twist Human Methylome Panel targets 3.98M CpG sites through 123 Mb of genomic content to target biologically relevant methylation markers. Expansive content makes this panel an ideal choice for investigators to explore the methylation fraction in a diverse range of applications from cancer metastasis, human development, and functional genomics.

The panel is optimized and validated for use with the Twist methylation detection system for a complete end-to-end workflow with industry leading performance. High capture efficiency increases the sensitivity of detection across the footprint of the epigenome while decreasing sequencing costs. The panel is ideal for screening cohort samples and differentially methylated region discovery.

Design and workflow efficiency

Optimized performance with an end to end workflow
Increased library complexity
Flexibility in adding spike-in content

Broader Coverage versus average microarray

Up to ~5.9x more coverage of all CpG sites
More coverage of CpG island bases
Based on updated reference databases

Advantage over microarrays

Higher calls at differentially methylated regions
More upstream and downstream coverage of adjacent CpG sites

Design and Workflow Efficiency

A complete solution that produces highly complex and uniform sequencing reads for methylation analysis. The end-to-end protocol achieves this by combining an innovative, enzymatic conversion process, optimized target enrichment workflow, and highly developed panel design process.

Broader Coverage versus Average Microarray

The Twist Human Methylome contains 3.98M CpG sites through the 123 Mb of genomic content target biologically relevant methylation markers. The panel is highly targeted to capture and detect the most current, annotated, and relevant CpG methylation regions in the genome. 84% (17,915,988) of the CpG islands in the genome are identified through enrichment with this panel. An additional 105,288,339 bases of subsequent shores, shelves, and open sea CpG’s and base pairs are covered. Coverage overlap determined by internal experimental data of coverage, including shadow coverage compared to array coverage data files.

Advantage Over Average Microarrays

A microarray is designed with static content that can hinder discovery of new epigenetic targets of interest. Current arrays on the market also have limited coverage of the methylome across the epigenome leaving a large proportion of Methylated CpG sites left unmeasured. The Twist Human Methylome targeted enrichment approach addresses these limitations with hybrid-capture panels that enable expanded content and single base resolution across the entire sequence read provided by NGS platforms.

Microarrays have inherent limitations on extreme ends of methylation detection attributed to high background noise at the low end and signal saturation at the high end. Because of the higher dynamic range provided by NGS, the Twist Human Methylome Panel can provide a more accurate calling of differentially methylated regions (DMRs) especially at both the low and high end of the methylation fraction.

Performance

The Twist Human Methylome Panel is designed and wet lab optimized to provide accurate, sensitive and high performance hybrid-capture efficiency. The panel achieves a depth of coverage of 90% of bases at 30x coverage with high probe specificities of 95% on-target rates. The panel also achieves a fold 80 of 1.54 providing high uniformity across the target region by decreasing the delta between the peaks and valleys of the base call pile ups. Library complexity was inversely represented by a <4% duplication rate at a high sequencing depth of 150x raw coverage. The overall panel capture efficiency metrics provide a high level of confidence in detection across the methylation fraction with minimized sequencing cost.