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(251r) A Microfluidic Device for Low-Input Methylomic Analysis Based on Reduced Representative Bisulfite Sequencing

Ma, S., Virginia Tech
Sun, C., Virginia Tech
Sun, Z., Virginia Tech
Murphy, T., Virginia Tech
Lu, C., Virginia Tech
Methylomic studies require substantial amounts of DNA samples and this restriction hinders applications involving scarce animal or patient samples with direct biomedical relevance. Here we report a microfluidics-based reduced representative bisulfite sequencing (RRBS) protocol that permits methylomic profiling with sub-1 ng starting DNA. We used diffusion-based reagent swapping for the multi-step treatment and purification of DNA on the microfluidic platform. Our device and protocol effectively minimized the loss of function for DNA to < 70% while achieving high bisulfite conversion within a 2 h period (in comparison to up to 96% DNA function loss in conventional bisulfite conversion protocol due to DNA hydrolysis under acidic condition). Using this technology, we studied DNA methylation in neurons and glia isolated from mouse cerebellum, revealing cell-type specific methylomic patterns and their influence on gene activities.