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Nat Commun. 2019 Feb 4;10(1):579. doi: 10.1038/s41467-019-08289-9.

Single-molecule sequencing detection of N6-methyladenine in microbial reference materials.

Author information

1
Department of Physiology and Biophysics, Weill Cornell Medicine, New York, 10065, NY, USA.
2
Tri-Institutional Training Program in Computational Biology and Medicine, New York, 10065, NY, USA.
3
US Food and Drug Administration, Silver Spring, 20993, MD, USA.
4
Department of Laboratory Medicine, University of California San Francisco, San Francisco, 94107, CA, USA.
5
UCSF-Abbott Viral Diagnostics and Discovery Center, San Francisco, 94107, CA, USA.
6
Department of Physiology and Biophysics, Weill Cornell Medicine, New York, 10065, NY, USA. chm2042@med.cornell.edu.
7
The HRH Prince Alwaleed Bin Talal Bin Abdulaziz Alsaud Institute for Computational Biomedicine, Weill Cornell Medicine, New York, 10021, NY, USA. chm2042@med.cornell.edu.
8
The Feil Family Brain and Mind Research Institute, Weill Cornell Medicine, New York, 10021, NY, USA. chm2042@med.cornell.edu.
9
The WorldQuant Initiative for Quantitative Prediction, Weill Cornell Medicine, New York, 10021, NY, USA. chm2042@med.cornell.edu.

Abstract

The DNA base modification N6-methyladenine (m6A) is involved in many pathways related to the survival of bacteria and their interactions with hosts. Nanopore sequencing offers a new, portable method to detect base modifications. Here, we show that a neural network can improve m6A detection at trained sequence contexts compared to previously published methods using deviations between measured and expected current values as each adenine travels through a pore. The model, implemented as the mCaller software package, can be extended to detect known or confirm suspected methyltransferase target motifs based on predictions of methylation at untrained contexts. We use PacBio, Oxford Nanopore, methylated DNA immunoprecipitation sequencing (MeDIP-seq), and whole-genome bisulfite sequencing data to generate and orthogonally validate methylomes for eight microbial reference species. These well-characterized microbial references can serve as controls in the development and evaluation of future methods for the identification of base modifications from single-molecule sequencing data.

PMID:
30718479
PMCID:
PMC6362088
DOI:
10.1038/s41467-019-08289-9
[Indexed for MEDLINE]
Free PMC Article

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