{"doi":"10.1007/978-1-4939-7231-9_8","title":"DNA Methylation Profiling Using Long-Read Single Molecule Real-Time Bisulfite Sequencing (SMRT-BS)","abstract":null,"journal":"Methods in Molecular Biology","year":2017,"id":590437,"datarank":1.109702820875774,"base_score":3.295836866004329,"endowment":3.295836866004329,"self_citation_contribution":0.4943755299006494,"citation_network_contribution":0.6153272909751246,"self_endowment_contribution":0.4943755299006494,"citer_contribution":0.6153272909751246,"corpus_percentile":null,"corpus_rank":null,"citation_count":26,"citer_count":26,"citers_with_citation_signal":21,"citers_with_endowment":21,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":235380,"name":"Stuart A. Scott","orcid":"0000-0001-5720-1864","position":1,"is_corresponding":false},{"id":942448,"name":"Yao Yang","orcid":"0000-0003-0784-8859","position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"DNA Methylation Profiling Using Long-Read Single Molecule Real-Time Bisulfite Sequencing (SMRT-BS)","abstract":"For the past two decades, bisulfite sequencing has been a widely used method for quantitative CpG methylation detection of genomic DNA. Coupled with PCR amplicon cloning, bisulfite Sanger sequencing allows for allele-specific CpG methylation assessment; however, its time-consuming protocol and inability to multiplex has recently been overcome by next-generation bisulfite sequencing techniques. Although high-throughput sequencing platforms have enabled greater accuracy in CpG methylation quantitation as a result of increased bisulfite sequencing depth, most common sequencing platforms generate reads that are similar in length to the typical bisulfite PCR size range (~300-500 bp). Using the Pacific Biosciences (PacBio) sequencing platform, we developed single molecule real-time bisulfite sequencing (SMRT-BS), which is an accurate targeted CpG methylation analysis method capable of a high degree of multiplexing and long read lengths. SMRT-BS is reproducible and was found to be concordant with other lower throughput quantitative CpG methylation methods. Moreover, the ability to sequence up to ~1.5-2.0 kb amplicons, when coupled with an optimized bisulfite-conversion protocol, allows for more thorough assessment of CpG islands and increases the capacity for studying the relationship between single nucleotide variants and allele-specific CpG methylation.","is_dataset_classified":null,"base_score":3.295836866004329,"endowment":3.295836866004329,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"28986786","pmcid":null,"openalex_id":"https://openalex.org/W2754684887","authors":[],"funders":[],"total_grants":0,"fwci":4.3502,"citation_percentile":0.94739857,"influential_citations":0,"citation_trend":[{"year":2018,"count":5},{"year":2019,"count":3},{"year":2020,"count":3},{"year":2021,"count":5},{"year":2022,"count":1},{"year":2023,"count":4},{"year":2024,"count":2},{"year":2025,"count":2},{"year":2026,"count":1}],"oa_status":"closed","license":"http://www.springer.com/tdm","oa_locations":[{"url":"http://link.springer.com/content/pdf/10.1007/978-1-4939-7231-9_8","host_type":"publisher"},{"url":"https://doi.org/10.1007/978-1-4939-7231-9_8","host_type":"book series"},{"url":"https://pubmed.ncbi.nlm.nih.gov/28986786","host_type":"repository"}],"fields_of_study":["Epigenetics and DNA Methylation","RNA modifications and cancer","Cancer-related gene regulation"],"mesh_terms":["Humans","Sulfites","Polymerase Chain Reaction","Sequence Analysis, DNA","CpG Islands","DNA Methylation","Nanotechnology","High-Throughput Nucleotide Sequencing"],"keywords":["Bisulfite","Bisulfite sequencing","Illumina Methylation Assay","Sanger sequencing","Methylated DNA immunoprecipitation","DNA methylation","CpG site","Methylation","Biology","Multiplex","Computational biology","Amplicon","Molecular biology","Genetics","DNA sequencing","DNA","Polymerase chain reaction","Gene","Gene expression","Cpg Islands","Pacbio Sequencing","Smrt Sequencing","Long-read Sequencing","Multiplex Dna Methylation Analysis"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Life below water"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-07-24T21:16:03.378147Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}