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

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 microflui...

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Detalles Bibliográficos
Autores principales: Budassi, Julia, Cho, NaHyun, Del Valle, Anthony, Sokolov, Jonathan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2023
Materias:
Research Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10482287/
https://www.ncbi.nlm.nih.gov/pubmed/37672538
http://dx.doi.org/10.1371/journal.pone.0250054
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author Budassi, Julia
Cho, NaHyun
Del Valle, Anthony
Sokolov, Jonathan
author_facet Budassi, Julia
Cho, NaHyun
Del Valle, Anthony
Sokolov, Jonathan
author_sort Budassi, Julia
collection PubMed
description 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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spelling pubmed-104822872023-09-07 Microfluidic delivery of cutting enzymes for fragmentation of surface-adsorbed DNA molecules Budassi, Julia Cho, NaHyun Del Valle, Anthony Sokolov, Jonathan PLoS One Research Article 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. Public Library of Science 2023-09-06 /pmc/articles/PMC10482287/ /pubmed/37672538 http://dx.doi.org/10.1371/journal.pone.0250054 Text en © 2023 Budassi et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Budassi, Julia
Cho, NaHyun
Del Valle, Anthony
Sokolov, Jonathan
Microfluidic delivery of cutting enzymes for fragmentation of surface-adsorbed DNA molecules
title Microfluidic delivery of cutting enzymes for fragmentation of surface-adsorbed DNA molecules
title_full Microfluidic delivery of cutting enzymes for fragmentation of surface-adsorbed DNA molecules
title_fullStr Microfluidic delivery of cutting enzymes for fragmentation of surface-adsorbed DNA molecules
title_full_unstemmed Microfluidic delivery of cutting enzymes for fragmentation of surface-adsorbed DNA molecules
title_short Microfluidic delivery of cutting enzymes for fragmentation of surface-adsorbed DNA molecules
title_sort microfluidic delivery of cutting enzymes for fragmentation of surface-adsorbed dna molecules
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10482287/
https://www.ncbi.nlm.nih.gov/pubmed/37672538
http://dx.doi.org/10.1371/journal.pone.0250054
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