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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...

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Autores principales: Lee, Jong-Han, Park, Yongjung, Choi, Jong Rak, Lee, Eun Kyung, Kim, Hyon-Suk
Formato: Texto
Lenguaje:English
Publicado: Yonsei University College of Medicine 2010
Materias:
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.
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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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