Microfluidic delivery of cutting enzymes for fragmentation of surface-adsorbed DNA molecules

Author:

Budassi Julia,Cho NaHyun,Del Valle AnthonyORCID,Sokolov JonathanORCID

Abstract

We describe a method for fragmenting, in-situ, surface-adsorbed and immobilized DNAs on polymethylmethacrylate(PMMA)-coated silicon substrates using microfluidic delivery of the cutting enzyme DNase I. Soft lithography is used to produce silicone elastomer (Sylgard 184) gratings which form microfluidic channels for delivery of the enzyme. Bovine serum albumin (BSA) is used to reduce DNase I adsorption to the walls of the microchannels and enable diffusion of the cutting enzyme to a distance of 10mm. Due to the DNAs being immobilized, the fragment order is maintained on the surface. Possible methods of preserving the order for application to sequencing are discussed.

Funder

National Science Foundation

Office of Science

Publisher

Public Library of Science (PLoS)

Subject

Multidisciplinary

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