(346c) DNA Sample Preparation for Long-Read Genomics Using Microfluidics
AIChE Annual Meeting
2019
2019 AIChE Annual Meeting
Food, Pharmaceutical & Bioengineering Division
Micro- and Nano-Scale Technologies in Life Sciences II
Tuesday, November 12, 2019 - 1:06pm to 1:24pm
We have developed a poly(dimethylsiloxane) device, inspired by 3D cell culture, to extract high molecular weight DNA from cells in a gel matrix. Rapid DNA purification is achieved by continuous flow in communicating microfluidic channels, which facilitate subsequent electrophoretic extraction of the long DNA out of the miniaturized gel. The disposable PDMS device produces 10 ng DNA from human breast cancer cells at greater than 80% recovery rate. Molecular weight distribution of the device-extracted DNA sample is obtained by molecular combing in microchannels. More than 80% of the characterized DNA molecules are longer than 100 kilobase pairs in size, with the longest molecule extending up to 4 megabase pairs. Our microfluidic cells-to-long DNA process, which is completed in 4 hours, significantly reduces the time, labor and reagent costs associated with long DNA sample preparation. The single-layer microfluidic outlay of the device holds potential for direct integration with genomics chips to obtain a total analysis system.