biostudies-literatureTham CYMOH | National Medical Research Council281https://www.ebi.ac.uk/biostudies/studies/S-EPMC9845338biostudies-literatureUnknown14(1)Telomeres are specialized nucleoprotein structures at the ends of linear chromosomes. The progressive shortening of steady-state telomere length in normal human somatic cells is a promising biomarker for age-associated diseases. However, there remain substantial challenges in quantifying telomere length due to the lack of high-throughput method with nucleotide resolution for individual telomere. Here, we describe a workflow to capture telomeres using newly designed telobaits in human culture cell lines as well as clinical patient samples and measure their length accurately at nucleotide resolution using single-molecule real-time (SMRT) sequencing. Our results also reveal the extreme heterogeneity of telomeric variant sequences (TVSs) that are dispersed throughout the telomere repeat regionNature communicationsHigh-throughput telomere length measurement at nucleotide resolution using the PacBio high fidelity sequencing platform.PMC9845338NMRC/CIRG/1481/2017MOH-000153Xu LLiu JKoh JYPHwang WYKYan TKoh ASZhang SLee GSTeh BTOsato MLi SHong ZSong HWPoon LCai YTham CYTan PRamlee MKTeoh VSIfalseHigh-throughput telomere length measurement at nucleotide resolution using the PacBio high fidelity sequencing platform.Telomeres are specialized nucleoprotein structures at the ends of linear chromosomes. The progressive shortening of steady-state telomere length in normal human somatic cells is a promising biomarker for age-associated diseases. However, there remain substantial challenges in quantifying telomere length due to the lack of high-throughput method with nucleotide resolution for individual telomere. Here, we describe a workflow to capture telomeres using newly designed telobaits in human culture cell lines as well as clinical patient samples and measure their length accurately at nucleotide resolution using single-molecule real-time (SMRT) sequencing. Our results also reveal the extreme heterogeneity of telomeric variant sequences (TVSs) that are dispersed throughout the telomere repeat region2023-01-01T00:00:00Z2023 Jan2026-06-03T18:30:27.576Z2025-04-19T12:37:57.216ZS-EPMC98453383665015510.1038/s41467-023-35823-7