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Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking

Background: Limited infrastructure is available to collect, store and transport venous blood in field epidemiological studies. Dried blood spot (DBS) is a robust potential alternative sample source for epidemiological studies & bio banking. A stable source of genomic DNA (gDNA) is required for l...

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Autores principales: Kumar, Abhinendra, Mhatre, Sharayu, Godbole, Sheela, Jha, Prabhat, Dikshit, Rajesh
Formato: Online Artículo Texto
Lenguaje:English
Publicado: F1000 Research Limited 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6883222/
https://www.ncbi.nlm.nih.gov/pubmed/31815249
http://dx.doi.org/10.12688/gatesopenres.12855.3
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author Kumar, Abhinendra
Mhatre, Sharayu
Godbole, Sheela
Jha, Prabhat
Dikshit, Rajesh
author_facet Kumar, Abhinendra
Mhatre, Sharayu
Godbole, Sheela
Jha, Prabhat
Dikshit, Rajesh
author_sort Kumar, Abhinendra
collection PubMed
description Background: Limited infrastructure is available to collect, store and transport venous blood in field epidemiological studies. Dried blood spot (DBS) is a robust potential alternative sample source for epidemiological studies & bio banking. A stable source of genomic DNA (gDNA) is required for long term storage in bio bank for its downstream applications. Our objective is to optimize the methods of gDNA extraction from stored DBS and with the aim of revealing its utility in large scale epidemiological studies.  Methods: The purpose of this study was to extract the maximum amount of gDNA from DBS on Whatman 903 protein saver card. gDNA was extracted through column  (Qiagen) & magnetic bead based (Invitrogen) methods. Quantification of extracted gDNA was performed with a spectrophotometer, fluorometer, and integrity analyzed by agarose gel electrophoresis.  Result: Large variation was observed in quantity & purity (260/280 ratio, 1.8-2.9) of the extracted gDNA. The intact gDNA bands on the electrophoresis gel reflect the robustness of DBS for gDNA even after prolonged storage time. The extracted gDNA amount 2.16 – 24 ng/µl is sufficient for its PCR based downstream application, but unfortunately it can’t be used for whole genome sequencing or genotyping from extracted gDNA. Sequencing or genotyping can be achieved by after increasing template copy number through whole genome amplification of extracted gDNA. The obtained results create a base for future research to develop high-throughput research and extraction methods from blood samples. Conclusion: The above results reveal, DBS can be utilized as a potential and robust sample source for bio banking in field epidemiological studies.
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spelling pubmed-68832222019-12-06 Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking Kumar, Abhinendra Mhatre, Sharayu Godbole, Sheela Jha, Prabhat Dikshit, Rajesh Gates Open Res Research Article Background: Limited infrastructure is available to collect, store and transport venous blood in field epidemiological studies. Dried blood spot (DBS) is a robust potential alternative sample source for epidemiological studies & bio banking. A stable source of genomic DNA (gDNA) is required for long term storage in bio bank for its downstream applications. Our objective is to optimize the methods of gDNA extraction from stored DBS and with the aim of revealing its utility in large scale epidemiological studies.  Methods: The purpose of this study was to extract the maximum amount of gDNA from DBS on Whatman 903 protein saver card. gDNA was extracted through column  (Qiagen) & magnetic bead based (Invitrogen) methods. Quantification of extracted gDNA was performed with a spectrophotometer, fluorometer, and integrity analyzed by agarose gel electrophoresis.  Result: Large variation was observed in quantity & purity (260/280 ratio, 1.8-2.9) of the extracted gDNA. The intact gDNA bands on the electrophoresis gel reflect the robustness of DBS for gDNA even after prolonged storage time. The extracted gDNA amount 2.16 – 24 ng/µl is sufficient for its PCR based downstream application, but unfortunately it can’t be used for whole genome sequencing or genotyping from extracted gDNA. Sequencing or genotyping can be achieved by after increasing template copy number through whole genome amplification of extracted gDNA. The obtained results create a base for future research to develop high-throughput research and extraction methods from blood samples. Conclusion: The above results reveal, DBS can be utilized as a potential and robust sample source for bio banking in field epidemiological studies. F1000 Research Limited 2019-11-14 /pmc/articles/PMC6883222/ /pubmed/31815249 http://dx.doi.org/10.12688/gatesopenres.12855.3 Text en Copyright: © 2019 Kumar A et al. http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Kumar, Abhinendra
Mhatre, Sharayu
Godbole, Sheela
Jha, Prabhat
Dikshit, Rajesh
Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking
title Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking
title_full Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking
title_fullStr Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking
title_full_unstemmed Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking
title_short Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking
title_sort optimization of extraction of genomic dna from archived dried blood spot (dbs): potential application in epidemiological research & bio banking
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6883222/
https://www.ncbi.nlm.nih.gov/pubmed/31815249
http://dx.doi.org/10.12688/gatesopenres.12855.3
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