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Comparisons of Three Automated Systems for Genomic DNA Extraction in a Clinical Diagnostic Laboratory
PURPOSE: The extraction of nucleic acid is initially a limiting step for successful molecular-based diagnostic workup. This study aims to compare the effectiveness of three automated DNA extraction systems for clinical laboratory use. MATERIALS AND METHODS: Venous blood samples from 22 healthy volun...
Autores principales: | , , , , |
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Formato: | Texto |
Lenguaje: | English |
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Yonsei University College of Medicine
2010
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2799962/ https://www.ncbi.nlm.nih.gov/pubmed/20046522 http://dx.doi.org/10.3349/ymj.2010.51.1.104 |
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author | Lee, Jong-Han Park, Yongjung Choi, Jong Rak Lee, Eun Kyung Kim, Hyon-Suk |
author_facet | Lee, Jong-Han Park, Yongjung Choi, Jong Rak Lee, Eun Kyung Kim, Hyon-Suk |
author_sort | Lee, Jong-Han |
collection | PubMed |
description | PURPOSE: The extraction of nucleic acid is initially a limiting step for successful molecular-based diagnostic workup. This study aims to compare the effectiveness of three automated DNA extraction systems for clinical laboratory use. MATERIALS AND METHODS: Venous blood samples from 22 healthy volunteers were analyzed using QIAamp® Blood Mini Kit (Qiagen), MagNA Pure LC Nucleic Acid Isolation Kit I (Roche), and Magtration-Magnazorb DNA common kit-200N (PSS). The concentration of extracted DNAs was measured by NanoDrop ND-1000 (PeqLab). Also, extracted DNAs were confirmed by applying in direct agarose gel electrophoresis and were amplified by polymerase chain reaction (PCR) for human beta-globin gene. RESULTS: The corrected concentrations of extracted DNAs were 25.42 ± 8.82 ng/µL (13.49-52.85 ng/µL) by QIAamp® Blood Mini Kit (Qiagen), and 22.65 ± 14.49 ng/µL (19.18-93.39 ng/µL) by MagNA Pure LC Nucleic Acid Isolation Kit I, and 22.35 ± 6.47 ng/µL (12.57-35.08 ng/µL) by Magtration-Magnazorb DNA common kit-200N (PSS). No statistically significant difference was noticed among the three commercial kits (p > 0.05). Only the mean value of DNA purity through PSS was slightly lower than others. All the extracted DNAs were successfully identified in direct agarose gel electrophoresis. And all the product of beta-globin gene PCR showed a reproducible pattern of bands. CONCLUSION: The effectiveness of the three automated extraction systems is of an equivalent level and good enough to produce reasonable results. Each laboratory could select the automated system according to its clinical and laboratory conditions. |
format | Text |
id | pubmed-2799962 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2010 |
publisher | Yonsei University College of Medicine |
record_format | MEDLINE/PubMed |
spelling | pubmed-27999622010-01-01 Comparisons of Three Automated Systems for Genomic DNA Extraction in a Clinical Diagnostic Laboratory Lee, Jong-Han Park, Yongjung Choi, Jong Rak Lee, Eun Kyung Kim, Hyon-Suk Yonsei Med J Original Article PURPOSE: The extraction of nucleic acid is initially a limiting step for successful molecular-based diagnostic workup. This study aims to compare the effectiveness of three automated DNA extraction systems for clinical laboratory use. MATERIALS AND METHODS: Venous blood samples from 22 healthy volunteers were analyzed using QIAamp® Blood Mini Kit (Qiagen), MagNA Pure LC Nucleic Acid Isolation Kit I (Roche), and Magtration-Magnazorb DNA common kit-200N (PSS). The concentration of extracted DNAs was measured by NanoDrop ND-1000 (PeqLab). Also, extracted DNAs were confirmed by applying in direct agarose gel electrophoresis and were amplified by polymerase chain reaction (PCR) for human beta-globin gene. RESULTS: The corrected concentrations of extracted DNAs were 25.42 ± 8.82 ng/µL (13.49-52.85 ng/µL) by QIAamp® Blood Mini Kit (Qiagen), and 22.65 ± 14.49 ng/µL (19.18-93.39 ng/µL) by MagNA Pure LC Nucleic Acid Isolation Kit I, and 22.35 ± 6.47 ng/µL (12.57-35.08 ng/µL) by Magtration-Magnazorb DNA common kit-200N (PSS). No statistically significant difference was noticed among the three commercial kits (p > 0.05). Only the mean value of DNA purity through PSS was slightly lower than others. All the extracted DNAs were successfully identified in direct agarose gel electrophoresis. And all the product of beta-globin gene PCR showed a reproducible pattern of bands. CONCLUSION: The effectiveness of the three automated extraction systems is of an equivalent level and good enough to produce reasonable results. Each laboratory could select the automated system according to its clinical and laboratory conditions. Yonsei University College of Medicine 2010-01-01 2009-12-29 /pmc/articles/PMC2799962/ /pubmed/20046522 http://dx.doi.org/10.3349/ymj.2010.51.1.104 Text en © Copyright: Yonsei University College of Medicine 2010 http://creativecommons.org/licenses/by-nc/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Original Article Lee, Jong-Han Park, Yongjung Choi, Jong Rak Lee, Eun Kyung Kim, Hyon-Suk Comparisons of Three Automated Systems for Genomic DNA Extraction in a Clinical Diagnostic Laboratory |
title | Comparisons of Three Automated Systems for Genomic DNA Extraction in a Clinical Diagnostic Laboratory |
title_full | Comparisons of Three Automated Systems for Genomic DNA Extraction in a Clinical Diagnostic Laboratory |
title_fullStr | Comparisons of Three Automated Systems for Genomic DNA Extraction in a Clinical Diagnostic Laboratory |
title_full_unstemmed | Comparisons of Three Automated Systems for Genomic DNA Extraction in a Clinical Diagnostic Laboratory |
title_short | Comparisons of Three Automated Systems for Genomic DNA Extraction in a Clinical Diagnostic Laboratory |
title_sort | comparisons of three automated systems for genomic dna extraction in a clinical diagnostic laboratory |
topic | Original Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2799962/ https://www.ncbi.nlm.nih.gov/pubmed/20046522 http://dx.doi.org/10.3349/ymj.2010.51.1.104 |
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