HAM-TBS: high-accuracy methylation measurements via targeted bisulfite sequencing

Epigenetics & Chromatin

卷:11

Susann Sauer¹  Nadine Provençal¹  Tobias Wiechmann¹  Simone Roeh¹  Maik Ködel¹  Elisabeth B. Binder¹ 

[1] Department of Translational Research in Psychiatry, Max Planck Institute of Psychiatry;

关键词: Targeted bisulfite sequencing;    DNA methylation;    Next-generation sequencing;    5-methylcytosine;    FKBP5;   

DOI  :  10.1186/s13072-018-0209-x

来源: DOAJ

【 摘 要 】

Abstract Background The ability to accurately and efficiently measure DNA methylation is critical to advance the understanding of this epigenetic mechanism and its contribution to common diseases. Here, we present a highly accurate method to measure methylation using bisulfite sequencing (termed HAM-TBS). This novel method is able to assess DNA methylation in multiple samples with high accuracy in a cost-effective manner. We developed this assay for the FKBP5 locus, an important gene in the regulation of the stress system and previously linked to stress-related disorders, but the method is applicable to any locus of interest. Results HAM-TBS enables multiplexed analyses of up to 96 samples and regions spanning 10 kb using the Illumina MiSeq. It incorporates a triplicate bisulfite conversion step, pooled target enrichment via PCR, PCR-free library preparation and a minimum coverage of 1000×. TBS was able to resolve DNA methylation levels with a mean accuracy of 0.72%. Using this method, we designed and validated a targeted panel to specifically assess regulatory regions within the FKBP5 locus that are not covered in commercially available DNA methylation arrays. Conclusions HAM-TBS represents a highly accurate, medium-throughput sequencing approach for robust detection of DNA methylation changes in specific target regions.

【 授权许可】

Unknown