Isolation and Quantification of Plasma Cell-Free DNA Using Different Manual and Automated Methods
Plasma cell-free DNA (cfDNA) originates from various tissues and cell types and can enable minimally invasive diagnosis, treatment and monitoring of cancer and other diseases. Proper extraction of cfDNA is critical to obtain optimal yields and purity. The goal of this study was to compare the perfor...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-10-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/12/10/2550 |
| _version_ | 1797473791503237120 |
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| author | Eleni Polatoglou Zsuzsanna Mayer Vida Ungerer Abel J. Bronkhorst Stefan Holdenrieder |
| author_facet | Eleni Polatoglou Zsuzsanna Mayer Vida Ungerer Abel J. Bronkhorst Stefan Holdenrieder |
| author_sort | Eleni Polatoglou |
| collection | DOAJ |
| description | Plasma cell-free DNA (cfDNA) originates from various tissues and cell types and can enable minimally invasive diagnosis, treatment and monitoring of cancer and other diseases. Proper extraction of cfDNA is critical to obtain optimal yields and purity. The goal of this study was to compare the performance of six commercial cfDNA kits to extract pure, high-quality cfDNA from human plasma samples and evaluate the quantity and size profiles of cfDNA extracts—among them, two spin-column based, three magnetic bead-based and two automatic magnetic bead-based methods. Significant differences were observed in the yield of DNA among the different extraction kits (up to 4.3 times), as measured by the Qubit Fluorometer and Bioanalyzer. All kits isolated mostly small fragments corresponding to mono-nucleosomal sizes. The highest yield and reproducibility were obtained by the manual QIAamp Circulating Nucleic Acid Kit and automated MagNA Pure Total NA Isolation Kit. The results highlight the importance of standardizing preanalytical conditions depending on the requirements of the downstream applications. |
| first_indexed | 2024-03-09T20:21:27Z |
| format | Article |
| id | doaj.art-82bfbdc74c564faaa070e829dff2e33b |
| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-03-09T20:21:27Z |
| publishDate | 2022-10-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-82bfbdc74c564faaa070e829dff2e33b2023-11-23T23:47:14ZengMDPI AGDiagnostics2075-44182022-10-011210255010.3390/diagnostics12102550Isolation and Quantification of Plasma Cell-Free DNA Using Different Manual and Automated MethodsEleni Polatoglou0Zsuzsanna Mayer1Vida Ungerer2Abel J. Bronkhorst3Stefan Holdenrieder4Munich Biomarker Research Center, Institute of Laboratory Medicine, German Heart Centre, Technical University Munich, Lazarettstraße 36, D-80636 Munich, GermanyMunich Biomarker Research Center, Institute of Laboratory Medicine, German Heart Centre, Technical University Munich, Lazarettstraße 36, D-80636 Munich, GermanyMunich Biomarker Research Center, Institute of Laboratory Medicine, German Heart Centre, Technical University Munich, Lazarettstraße 36, D-80636 Munich, GermanyMunich Biomarker Research Center, Institute of Laboratory Medicine, German Heart Centre, Technical University Munich, Lazarettstraße 36, D-80636 Munich, GermanyMunich Biomarker Research Center, Institute of Laboratory Medicine, German Heart Centre, Technical University Munich, Lazarettstraße 36, D-80636 Munich, GermanyPlasma cell-free DNA (cfDNA) originates from various tissues and cell types and can enable minimally invasive diagnosis, treatment and monitoring of cancer and other diseases. Proper extraction of cfDNA is critical to obtain optimal yields and purity. The goal of this study was to compare the performance of six commercial cfDNA kits to extract pure, high-quality cfDNA from human plasma samples and evaluate the quantity and size profiles of cfDNA extracts—among them, two spin-column based, three magnetic bead-based and two automatic magnetic bead-based methods. Significant differences were observed in the yield of DNA among the different extraction kits (up to 4.3 times), as measured by the Qubit Fluorometer and Bioanalyzer. All kits isolated mostly small fragments corresponding to mono-nucleosomal sizes. The highest yield and reproducibility were obtained by the manual QIAamp Circulating Nucleic Acid Kit and automated MagNA Pure Total NA Isolation Kit. The results highlight the importance of standardizing preanalytical conditions depending on the requirements of the downstream applications.https://www.mdpi.com/2075-4418/12/10/2550liquid biopsycell-free DNAplasmacell-free DNA isolation |
| spellingShingle | Eleni Polatoglou Zsuzsanna Mayer Vida Ungerer Abel J. Bronkhorst Stefan Holdenrieder Isolation and Quantification of Plasma Cell-Free DNA Using Different Manual and Automated Methods Diagnostics liquid biopsy cell-free DNA plasma cell-free DNA isolation |
| title | Isolation and Quantification of Plasma Cell-Free DNA Using Different Manual and Automated Methods |
| title_full | Isolation and Quantification of Plasma Cell-Free DNA Using Different Manual and Automated Methods |
| title_fullStr | Isolation and Quantification of Plasma Cell-Free DNA Using Different Manual and Automated Methods |
| title_full_unstemmed | Isolation and Quantification of Plasma Cell-Free DNA Using Different Manual and Automated Methods |
| title_short | Isolation and Quantification of Plasma Cell-Free DNA Using Different Manual and Automated Methods |
| title_sort | isolation and quantification of plasma cell free dna using different manual and automated methods |
| topic | liquid biopsy cell-free DNA plasma cell-free DNA isolation |
| url | https://www.mdpi.com/2075-4418/12/10/2550 |
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