Protocol for high-throughput DNA methylation profiling in rat tissues using automated reduced representation bisulfite sequencing

被引：0
|
作者：
Nair, Venugopalan D. [1 ]
Pincas, Hanna [1 ]
Amper, Mary Anne S. [1 ]
Ge, Yongchao [1 ]
Vasoya, Mital [1 ]
Raja, Archana Natarajan [2 ]
Walsh, Martin J. [3 ]
Sealfon, Stuart C. [1 ]
机构：
[1] Icahn Sch Med Mt Sinai, Dept Neurol, New York, NY 10029 USA
[2] Stanford Sch Med, Dept Med, Stanford, CA 94305 USA
[3] Icahn Sch Med Mt Sinai, Dept Pharmacol Sci, New York, NY 10029 USA
来源：
STAR PROTOCOLS | 2024年 / 5卷 / 02期
关键词：
Genomics; High-Throughput Screening; Molecular Biology;
D O I：
10.1016/j.xpro.2024.103007
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
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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页数：27
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