NGS Methylation Detection System

In partnership with New England Biolabs (NEB), Twist Bioscience offers a new methylation sequencing workflow that improves the quality of libraries and removes the need for damaging bisulfite treatment during prep. 
 
The workflow features enzymatic conversion of unmethylated cytosines (see figure) to identify sites of methyl-cytosine (5mC) and hydroxymethyl-cytosine (5hmC). 
 
Enzymatic conversion produces more intact libraries with better representation, and ultimately achieves more sensitive methylation detection. The library preparation system is suitable for whole genome sequencing and downstream enrichment with Twist Methylation Panels. 

EM-seq conversion involves a series of enzymatic steps to convert unmethylated cytosines into uracils. The final result is the same as conventional bisulfite conversion, making EM-seq compatible with existing analysis pipelines that use Bismark and bwa-meth. 

The Twist Fast Hybridization and Wash Kit provides optimal performance and maximum flexibility. Reagents and steps can be optimized to tune downstream sequencing metrics and to reduce hands-on time and pipette use. 

Designed with a highly sophisticated algorithm, Twist Custom Methylation Panels capture targets with exceptional efficiency across a wide range of target sizes. Together, these components ensure best-in-class hybrid capture of custom regions of interest.

Off-target capture frequently occurs in targeted methylation sequencing workflows because the conversion process reduces the sequence complexity of the sample during library preparation. 

To maximize the capture of post-conversion samples, the Twist Methylation Detection System features a proprietary blocker designed specifically for methylation detection. Called Methylation Enhancer, this reagent reduces off-target capture by as much as half.  
 

Methylation levels vary substantially across the human genome, and differentially methylated regions (DMRs) can be used to identify certain cancers. 

To test the effects of methylation levels on the performance of the Twist Methylation Detection System, libraries of varying methylation levels (0-100% methylation) were generated by combining hypo- and hypermethylated genomic DNA in defined ratios. This analysis showed minimal effects of methylation level on final sequencing metrics.

The Twist Methylation Detection System also captures hypo- and hypermethylated regions with high sensitivity.