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Background: DNA methylation is commonly measured using bisulfite sequencing (BS-seq). The quality of a BS-seq library is measured by its bisulfite conversion efficiency. Libraries with low conversion rates are typically excluded from analysis resulting in reduced coverage and increased costs.
Results: We have developed a probabilistic method and software, LuxRep, that implements a general linear model and simultaneously accounts for technical replicates (libraries from the same biological sample) from different bisulfite-converted DNA libraries. Using simulations and actual DNA methylation data, we show that including technical replicates with low bisulfite conversion rates generates more accurate estimates of methylation levels and differentially methylated sites. Moreover, using variational inference speeds up computation time necessary for whole genome analysis.
Conclusions: In this work we show that taking into account technical replicates (i.e. libraries) of BS-seq data of varying bisulfite conversion rates, with their corresponding experimental parameters, improves methylation level estimation and differential methylation detection.
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http://dx.doi.org/10.1186/s12859-021-04546-1 | DOI Listing |
Nat Commun
August 2025
State Key Laboratory of Metabolism and Regulation in Complex Organisms, College of Chemistry and Molecular Sciences, Taikang Center for Life and Medical Sciences, Wuhan University, Wuhan, China.
Extracellular vesicles (EVs) are promising biomarkers for cancer diagnosis and prognosis due to their ability to carry specific biomolecular cargo, including DNA. However, the clinical utility of DNA methylation-based liquid biopsies using EV-DNA remains underexplored. The low quantity and relatively long length of EV-DNA complicate whole-genome methylation profiling.
View Article and Find Full Text PDFInt J Mol Sci
August 2025
Department of Biology, Memorial University of Newfoundland, St. John's, NL A1C 5S7, Canada.
The cytosine methylation status of symmetric and asymmetric sites of promoters of the genes encoding the membrane-bound subunits C and D of succinate dehydrogenase (SDH) was assessed during the germination of maize ( L.) seeds, when the stored lipids were utilized and the glyoxylate cycle produced succinate. The results of bisulfite sequencing of the promoters of genes in maize scutella allowed us to determine the cytosine methylation status in the CG, CNG, and CNN sites.
View Article and Find Full Text PDFFEBS Open Bio
August 2025
GenPhySE, Université de Toulouse, INRAE, ENVT, Castanet-Tolosan, France.
Genome methylation represents an important source of regulation of gene expression. To date, custom molecular tools for studying targeted regions of the genome are restricted to several megabases. We developed a protocol to epigenotype differentially methylated CpGs in specific regions of the genome.
View Article and Find Full Text PDFEpigenomics
September 2025
Murdoch Children's Research Institute, Royal Children's Hospital, Parkville, VIC, Australia.
Aim: This study compared methylation-specific quantitative melt analysis of FMR1 and SNRPN methylation (mDNA) using automated bisulfite conversion by the magnetic-bead-based IsoPure and column-based QIAcube HT systems.
Methods: Two bisulfite conversion methods were assessed on 3.2 mm punches from the same archival blood spots stored at room temperature for >10 years of individuals with FMR1 premutation ( = 20), fragile X syndrome (FXS, = 20), or chromosome 15 imprinting disorders ( = 50) and freshly made blood spots from 184 newborns from the general population.
PLoS One
August 2025
Department of Pediatrics, Faculty of Medicine, Center of Excellence for Medical Genomics, Chulalongkorn University, Bangkok, Thailand.
DNA methylation, a key epigenetic modification, regulates gene expression and diverse cellular functions. Bisulfite sequencing (BS) remains the gold standard for methylation detection, while PacBio HiFi sequencing enables direct detection without chemical conversion. Although both technologies are increasingly used, few studies have directly compared their concordance, particularly in clinically relevant settings such as Down syndrome (DS).
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