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

被引：0
|
作者：
Budassi, Julia [1 ]
Cho, Nahyun [1 ]
Del Valle, Anthony [2 ]
Sokolov, Jonathan [1 ]
机构：
[1] SUNY Stony Brook, Dept Mat Sci & Chem Engn, Stony Brook, NY 11974 USA
[2] SUNY Stony Brook, Dept Phys & Astron, Stony Brook, NY USA
来源：
PLOS ONE | 2023年 / 18卷 / 09期
关键词：
POLYMERASE-CHAIN-REACTION; LIBRARY CONSTRUCTION; POLY(DIMETHYLSILOXANE); PDMS; DEVICES; GENOME; CHIP; ULTRAVIOLET; CONTIGUITY; ABSORPTION;
D O I：
10.1371/journal.pone.0250054
中图分类号：
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号：
07 ; 0710 ; 09 ;
摘要：
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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页数：14
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