Human Methylation Whole-Genome Sequencing (Cloudbreak™)

Cloudbreak chemistry was used to perform human methylation whole-genome sequencing (WGS) of reference genomic DNA (gDNA), preserving the methylation status of cytosine residues.

Methods

NA12878 was mixed with methylated pUC19 and unmethylated Lambda phage DNA, then sheared to a fragment size of ~250 bp. Two replicates of 200 ng sheared DNA were input for the NEBNext Enzymatic Methyl-seq Kit, which was paired with the NEBNext Multiplex Oligos for Enzymatic Methyl-seq (Unique Dual Index Primer Pairs). Samples were amplified, circularized with the Adept™ Library Compatibility Kit v1.1, and pooled equally. The AVITI™ System performed a 2 x 150 run and Bases2Fastq generated the FASTQ files.