The Accel-NGS Methyl-Seq DNA library kit from Swift Biosciences is optimised for methylation sequencing methods, which include whole genome bisulphite sequencing (WGBS), reduced representation bisulphite sequencing (RRBS) and targeted bisulphite sequencing, through hybridisation capture methods. By employing a post-bisulphite library preparation workflow to maximise DNA recovery, the Accel-NGS Methyl-Seq DNA library kit is suitable for even low input quantities such as liquid biopsy from cell free DNA (cfDNA) for oncology studies. In addition to this, sequence independent adaptor ligation avoids bias and allows coverage of AT-rich sequences produced by bisulphite conversion.
Benefits of using the Accel-NGS Methyl-Seq DNA library kit
• High recovery of input DNA
• Low bias library preparation
• Simple, 2-hour protocol
• Inputs from 100 pg to 100 ng
• Minimal PCR cycles required
• Suitable for ancient, uracil-containing DNA samples
• Reveals the complete methylome
• Reduced amplification errors
The Accel-NGS Methyl-Seq DNA library prep kit utilises Adaptase® technology for capturing single-stranded DNA molecules in an unbiased manner. The Accel-NGS Methyl-Seq Kit is also compatible with bisulphite-converted DNA samples enriched by ChIP or other methods.
Accel-NGS Methyl-Seq DNA library kit workflow comparison to traditional & random priming methods
For both Accel-NGS Methyl-Seq and random priming kits, bisulfite conversion is performed prior to library construction. With the traditional library kit, bisulfite conversion is performed on the completed library. The lightning bolts represent bisulfite-induced fragmentation, NGS adapters are depicted in green and blue, and non-uracil containing library products are shown in yellow.
Coverage of unique CpX and CpG dinucleotide sequences
(A, B) To assess methylome-specific coverage, 17.5 million unique CpX (CpG + CpH; where H is A, T, or C) dinucleotides were assessed from the Arabidopsis TAIR10 reference, including both completeness of coverage (A) and uniformity of coverage (B). (C) To assess methylome-specific coverage of the human genome, 28.1 million unique CpG dinucleotides were assessed for both completeness of coverage and coverage uniformity at 1X and 5X (where the average depth was 8.9X).
Material available for download
Application note - NGS library preparation produces balance, comprehensive methylome coverage from low input quantities
Accel NGS Methyl-Seq DNA-library kit datasheet
Accel NGS Methyl-Seq DNA-library kit protocol
DNA Input quantification assay
Accel-NGS 1S Plus & Methyl-Seq tail trimming technical note
Webinar: Lowering the limits for epigenetic methylation analysis
Video: Methyl-Seq DNA, sequence the 5th base of DNA
Swift product and indexing selection guide