Electro-DBS : A Simple Method to Rapidly Extract Genomic DNA from Dried Blood Spots
Dried blood spots (DBSs) have been used for more than 50 years and are used as a method of sample collection for pharmacological testing, genetic analyses, monitoring of viral infections, and more. Protocols for nucleic acid extraction from DBSs involve several steps that require specialized equipment and can be lengthy. Thus, we sought to explore ways to reduce the analytical burden of DBSs. We developed a DBS extraction method that uses the synergistic action of electrophoretic and diffusive transport mechanisms to extract genomic DNA (gDNA) through the DBS matrix. This method (which we termed "Electro-DBS") reduces the time of extraction from 40 min to 5 min and removes the need for heat, shaking, and DNA purification steps while maintaining gDNA quality and yield. We found that the electrophoretic transport speeds up gDNA elution, allowing for the diffusive transport of polymerase chain reaction (PCR) inhibitors to be minimized due to a reduced elution time. Overall, this work added mechanistic insight into the extraction of DNA from DBSs and developed a method that was ideal for point of care and automation.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2022 |
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| Erschienen: |
2022 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:94 |
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| Enthalten in: |
Analytical chemistry - 94(2022), 39 vom: 04. Okt., Seite 13404-13412 |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Lee, Kiara [VerfasserIn] |
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| Links: |
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| Themen: |
9007-49-2 |
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| Anmerkungen: |
Date Completed 05.10.2022 Date Revised 11.11.2022 published: Print-Electronic Citation Status MEDLINE |
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| doi: |
10.1021/acs.analchem.2c02021 |
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| funding: |
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| Förderinstitution / Projekttitel: |
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NLM345613589 |
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| 520 | |a Dried blood spots (DBSs) have been used for more than 50 years and are used as a method of sample collection for pharmacological testing, genetic analyses, monitoring of viral infections, and more. Protocols for nucleic acid extraction from DBSs involve several steps that require specialized equipment and can be lengthy. Thus, we sought to explore ways to reduce the analytical burden of DBSs. We developed a DBS extraction method that uses the synergistic action of electrophoretic and diffusive transport mechanisms to extract genomic DNA (gDNA) through the DBS matrix. This method (which we termed "Electro-DBS") reduces the time of extraction from 40 min to 5 min and removes the need for heat, shaking, and DNA purification steps while maintaining gDNA quality and yield. We found that the electrophoretic transport speeds up gDNA elution, allowing for the diffusive transport of polymerase chain reaction (PCR) inhibitors to be minimized due to a reduced elution time. Overall, this work added mechanistic insight into the extraction of DNA from DBSs and developed a method that was ideal for point of care and automation | ||
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