1. Simultaneous sequencing of genetic and epigenetic bases in DNA
- Author
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Jens Füllgrabe, Walraj S. Gosal, Páidí Creed, Sidong Liu, Casper K. Lumby, David J. Morley, Tobias W. B. Ost, Albert J. Vilella, Shirong Yu, Helen Bignell, Philippa Burns, Tom Charlesworth, Beiyuan Fu, Howerd Fordham, Nicolas J. Harding, Olga Gandelman, Paula Golder, Christopher Hodson, Mengjie Li, Marjana Lila, Yang Liu, Joanne Mason, Jason Mellad, Jack M. Monahan, Oliver Nentwich, Alexandra Palmer, Michael Steward, Minna Taipale, Audrey Vandomme, Rita Santo San-Bento, Ankita Singhal, Julia Vivian, Natalia Wójtowicz, Nathan Williams, Nicolas J. Walker, Nicola C. H. Wong, Gary N. Yalloway, Joanna D. Holbrook, and Shankar Balasubramanian
- Subjects
Biomedical Engineering ,Molecular Medicine ,Bioengineering ,Applied Microbiology and Biotechnology ,Biotechnology - Abstract
DNA comprises molecular information stored in genetic and epigenetic bases, both of which are vital to our understanding of biology. Most DNA sequencing approaches address either genetics or epigenetics and thus capture incomplete information. Methods widely used to detect epigenetic DNA bases fail to capture common C-to-T mutations or distinguish 5-methylcytosine from 5-hydroxymethylcytosine. We present a single base-resolution sequencing methodology that sequences complete genetics and the two most common cytosine modifications in a single workflow. DNA is copied and bases are enzymatically converted. Coupled decoding of bases across the original and copy strand provides a phased digital readout. Methods are demonstrated on human genomic DNA and cell-free DNA from a blood sample of a patient with cancer. The approach is accurate, requires low DNA input and has a simple workflow and analysis pipeline. Simultaneous, phased reading of genetic and epigenetic bases provides a more complete picture of the information stored in genomes and has applications throughout biomedicine.
- Published
- 2023
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