Protocol for High-Throughput DNA Methylation Profiling in Rat Tissues Using Automated Reduced Representation Bisulfite Sequencing

Venugopalan D. Nair,Hanna Pincas,Mary Anne S. Amper,Yongchao Ge,Mital Vasoya,Archana Natarajan Raja,Martin J. Walsh,Stuart C. Sealfon

STAR PROTOCOLS（2024）

Icahn Sch Med Mt Sinai | Stanford Sch Med

Cited 0|Views25

Abstract

Although reduced representation bisulfite sequencing (RRBS) measures DNA methylation (DNAme) across CpG-rich genomic regions with high sensitivity, the assay can be time-consuming and prone to batch effects. Here, we present a high -throughput, automated RRBS protocol starting with DNA extraction from frozen rat tissues. We describe steps for RRBS library preparation, library quality control, and sequencing. We also detail an optimized pipeline for sequencing data processing. This protocol has been applied successfully to DNAme profiling across multiple rat tissues. For complete details on the use and execution of this protocol, please refer to Nair et al. 1

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Genomics,High-Throughput Screening,Molecular Biology

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