ArticleNucleic acids research2001
Bisulfite genomic sequencing: systematic investigation of critical experimental parameters.
Article in Nucleic acids research, 2001. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 326 papers, 2 of them syntheses that pooled it.
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The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
326 citing papers in PubMed, 2 syntheses or guidelines pooled it, 790 citations in OpenAlex.
- Early diagnosis of ovarian cancer based on methylation profiles in peripheral blood cell-free DNA: a systematic review.Clinical epigenetics · 2023Pooled it
- Guidelines for pre-analytical conditions for assessing the methylation of circulating cell-free DNA.Clinical epigenetics · 2021Pooled it
- LINE-1 CpG methylation in canine blood and cortical DNA tracks epigenetic changes with age.GeroScience · 2026Article
- A novel highly specific qPCR assay for distinguishing HBV cccDNA from rcDNA following bisulfite pretreatment.Journal of virology · 2026Article
- Article
- Machine learning-assisted analysis of saliva-containing mixtures using a saliva-specific multi-CpG-microhaplotype marker system.International journal of legal medicine · 2026Article
- CRISPR technologies for detecting DNA and RNA methylation: Mechanisms, platforms, and translational opportunities.Bioengineering & translational medicine · 2026Review
- Comprehensive benchmarking of tools for nanopore-based detection of DNA methylation.Nature communications · 2026Article
- CMA-Nano: A DNA Methylation Detection Method for Nanopore Sequencing Data Based on a Cross-Modal Attention Mechanism.ACS omega · 2026Article
- Epigenetic age acceleration in adolescence: cross-sectional associations with dietary intake and prospective associations with cardiometabolic risk indicators in a Mexico City cohort.Nutrition, metabolism, and cardiovascular diseases : NMCD · 2026Article
- Establishment and Performance Evaluation of a Multiplexed TET2-APOBEC-Mediated cfDNA Methylation Detection Workflow Using qPCR and dPCR Readouts.Journal of personalized medicine · 2026Article
- Comparative validation of oxidative bisulfite sequencing (oxBS) and chemical-assisted pyridine borane sequencing (CAPS) protocols for locus-specific 5-hydroxymethylcytosine quantification.Epigenetics & chromatin · 2026Article
- The effects of ingested cellulose nanomaterials on DNA methylation in intestinal cells.Particle and fibre toxicology · 2026Article
- Brain tumor detection based on transcription regulation features identified from public cerebrospinal fluid cell-free DNA sequencing data.Translational cancer research · 2026Article
- Sequencing DNA methylation and hydroxymethylation at co-occurring chromatin features.Nature communications · 2026Article
- Haplotype-resolved DNA methylation at theNPJ dementia · 2026Article
- Next generation sequencing and beyond: a review of genomic sequencing methods.Functional & integrative genomics · 2025Article
- Ultra-mild bisulfite outperforms existing methods for 5-methylcytosine detection with low input DNA.Nature communications · 2025Article
- Computational Workflow for Genome-Wide DNA Methylation Profiling and Differential Methylation Analysis.Bio-protocol · 2025Article
- Adolescent dietary patterns and methyl-donor nutrient intakes in relation to blood leukocyte DNA methylation of circadian genes.Chronobiology international · 2025Article
266 more citing papers are in PubMed but not listed here.
Corrections and comments
PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.
Authors and funding
3 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Bisulfite genomic sequencing is the method of choice for the generation of methylation maps with single-base resolution. The method is based on the selective deamination of cytosine to uracil by treatment with bisulfite and the sequencing of subsequently generated PCR products. In contrast to cytosine, 5-methylcytosine does not react with bisulfite and can therefore be distinguished. In order to investigate the potential for optimization of the method and to determine the critical experimental parameters, we determined the influence of incubation time and incubation temperature on the deamination efficiency and measured the degree of DNA degradation during the bisulfite treatment. We found that maximum conversion rates of cytosine occurred at 55 degrees C (4-18 h) and 95 degrees C (1 h). Under these conditions at least 84-96% of the DNA is degraded. To study the impact of primer selection, homologous DNA templates were constructed possessing cytosine-containing and cytosine-free primer binding sites, respectively. The recognition rates for cytosine (>/=97%) and 5-methylcytosine (>/=94%) were found to be identical for both templates.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.