Microfluidic delivery of cutting enzymes for fragmentation of surface-adsorbed DNA molecules.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 37672538.
- Also identified by DOI 10.1371/journal.pone.0250054 and PMC identifier 10482287.
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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.
Medical subject headings
- Microfluidics
- DNA