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Digital DNA microarray generation on glass substrates
In this work we show how DNA microarrays can be produced batch wise on standard microscope slides in a fast, easy, reliable and cost-efficient way. Contrary to classical microarray generation, the microarrays are generated via digital solid phase PCR. We have developed a cavity-chip system made of a...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group UK
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7113318/ https://www.ncbi.nlm.nih.gov/pubmed/32238843 http://dx.doi.org/10.1038/s41598-020-62404-1 |
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author | Wöhrle, Johannes Krämer, Stefan D. Meyer, Philipp A. Rath, Christin Hügle, Matthias Urban, Gerald A. Roth, Günter |
author_facet | Wöhrle, Johannes Krämer, Stefan D. Meyer, Philipp A. Rath, Christin Hügle, Matthias Urban, Gerald A. Roth, Günter |
author_sort | Wöhrle, Johannes |
collection | PubMed |
description | In this work we show how DNA microarrays can be produced batch wise on standard microscope slides in a fast, easy, reliable and cost-efficient way. Contrary to classical microarray generation, the microarrays are generated via digital solid phase PCR. We have developed a cavity-chip system made of a PDMS/aluminum composite which allows such a solid phase PCR in a scalable and easy to handle manner. For the proof of concept, a DNA pool composed of two different DNA species was used to show that digital PCR is possible in our chips. In addition, we demonstrate that DNA microarray generation can be realized with different laboratory equipment (slide cycler, manually in water baths and with an automated cartridge system). We generated multiple microarrays and analyzed over 13,000 different monoclonal DNA spots to show that there is no significant difference between the used equipment. To show the scalability of our system we also varied the size and number of the cavities located in the array region up to more than 30,000 cavities with a volume of less than 60 pL per cavity. With this method, we present a revolutionary tool for novel DNA microarrays. Together with new established label-free measurement systems, our technology has the potential to give DNA microarray applications a new boost. |
format | Online Article Text |
id | pubmed-7113318 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-71133182020-04-06 Digital DNA microarray generation on glass substrates Wöhrle, Johannes Krämer, Stefan D. Meyer, Philipp A. Rath, Christin Hügle, Matthias Urban, Gerald A. Roth, Günter Sci Rep Article In this work we show how DNA microarrays can be produced batch wise on standard microscope slides in a fast, easy, reliable and cost-efficient way. Contrary to classical microarray generation, the microarrays are generated via digital solid phase PCR. We have developed a cavity-chip system made of a PDMS/aluminum composite which allows such a solid phase PCR in a scalable and easy to handle manner. For the proof of concept, a DNA pool composed of two different DNA species was used to show that digital PCR is possible in our chips. In addition, we demonstrate that DNA microarray generation can be realized with different laboratory equipment (slide cycler, manually in water baths and with an automated cartridge system). We generated multiple microarrays and analyzed over 13,000 different monoclonal DNA spots to show that there is no significant difference between the used equipment. To show the scalability of our system we also varied the size and number of the cavities located in the array region up to more than 30,000 cavities with a volume of less than 60 pL per cavity. With this method, we present a revolutionary tool for novel DNA microarrays. Together with new established label-free measurement systems, our technology has the potential to give DNA microarray applications a new boost. Nature Publishing Group UK 2020-04-01 /pmc/articles/PMC7113318/ /pubmed/32238843 http://dx.doi.org/10.1038/s41598-020-62404-1 Text en © The Author(s) 2020 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. |
spellingShingle | Article Wöhrle, Johannes Krämer, Stefan D. Meyer, Philipp A. Rath, Christin Hügle, Matthias Urban, Gerald A. Roth, Günter Digital DNA microarray generation on glass substrates |
title | Digital DNA microarray generation on glass substrates |
title_full | Digital DNA microarray generation on glass substrates |
title_fullStr | Digital DNA microarray generation on glass substrates |
title_full_unstemmed | Digital DNA microarray generation on glass substrates |
title_short | Digital DNA microarray generation on glass substrates |
title_sort | digital dna microarray generation on glass substrates |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7113318/ https://www.ncbi.nlm.nih.gov/pubmed/32238843 http://dx.doi.org/10.1038/s41598-020-62404-1 |
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