Enzymatic Methyl-Seq: Cytosine Methylation Detection Using Picograms Of Dna.

Cytosine methylation (5mC) is an important regulator of gene expression. Bisulfite sequencing is most commonly used to study 5mCs but it is frequently associated with DNA damage and fragmentation. This can prove problematic when using low amounts of DNA as the damage results in uneven genome coverage as well as biased sequencing data. Here we show data generated using NEBNext® Enzymatic Methyl-Seq (EM-Seq™), an enzymatic alternative to bisulfite conversion. The method has been optimized for low input DNAs ranging from 10 ng to 100 pg. NA12878 DNA was used to demonstrate that the modifications to the protocol permitted accurate representation of global CpG, CHG and CHH methylation. These libraries also displayed similar basic metrics to standard input EM-seq libraries. The GC bias plot was even and there was high CpG coverage with strong correlation between replicates. In addition, the possibility of introducing unique molecular identifiers into this system would be useful for many researchers who use DNA originating from single cells, plasma (cfDNA) and formalin fixed paraffin embedded (FFPE) sections. This low input method is robust and reproducible and gives researchers a new tool to tackle studies where DNA has traditionally been limiting. Citation Format: Louise Williams, V. K. Chaithanya Ponnaluri, Vaishnavi Panchapakesa, Romualdas Vaisvila, Matthew A. Campbell, Bradley W. Langhorst, Eileen T. Dimalanta, Theodore B. Davis. Enzymatic methyl-seq: Cytosine methylation detection using picograms of DNA [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2101.