479013 A Microfluidic Device for Low-Input Methylomic Analysis Based on Reduced Representative Bisulfite Sequencing

Monday, November 14, 2016
Embarcadero (Parc 55 San Francisco)
Sai Ma1, Chen Sun1, Zhixiong Sun2, Travis Murphy3, Hehuang Xie2 and Chang Lu4, (1)School of Biomedical Engineering and Sciences, Virginia Tech, Blacksburg, VA, (2)Virginia Bioinformatics Institute, Virginia Tech, Blacksburg, VA, Virginia Tech, Blacksburg, VA, (3)Chemical Engineering, Virginia Tech, Blacksburg, VA, Virginia Tech, Blacksburg, VA, (4)Chemical Engineering, Virginia Tech, Blacksburg, VA

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.

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See more of this Session: Poster Session: AES
See more of this Group/Topical: 2016 Annual Meeting of the AES Electrophoresis Society