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

Julia Budassi; Na Hyun Cho; Anthony Del Valle; Jonathan Sokolov

doi:10.1371/journal.pone.0250054

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

Julia Budassi
, Na Hyun Cho
, Anthony Del Valle
, Jonathan Sokolov

Materials Science and Engineering

Stony Brook University

Research output: Contribution to journal › Article › peer-review

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.

Original language	English
Article number	e0250054
Journal	PLoS ONE
Volume	18
Issue number	9 September
DOIs	https://doi.org/10.1371/journal.pone.0250054
State	Published - Sep 2023

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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.",

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AB - 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.

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

Abstract

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