Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking [version 3; peer review: 2 approved]
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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doaj-625043b2cf894169a10156fd09d5be9c2020-11-25T03:44:22ZengF1000 Research LtdGates Open Research2572-47542019-11-01210.12688/gatesopenres.12855.314229Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking [version 3; peer review: 2 approved]Abhinendra Kumar0Sharayu Mhatre1Sheela Godbole2Prabhat Jha3Rajesh Dikshit4Centre for Cancer Epidemiology, Tata Memorial Centre, Mumbai, Maharashtra, 410210, IndiaCentre for Cancer Epidemiology, Tata Memorial Centre, Mumbai, Maharashtra, 410210, IndiaDepartment of Biostatistics and Epidemiology, National AIDS Research Institute, Pune, Maharashtra, 411026, IndiaLi Ka Shing Knowledge Institiute, St Michael's Hospital, Center for Global Health Research, Toronto, ON, CanadaCentre for Cancer Epidemiology, Tata Memorial Centre, Mumbai, Maharashtra, 410210, IndiaBackground: 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.https://gatesopenresearch.org/articles/2-57/v3 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Abhinendra Kumar Sharayu Mhatre Sheela Godbole Prabhat Jha Rajesh Dikshit |
spellingShingle |
Abhinendra Kumar Sharayu Mhatre Sheela Godbole Prabhat Jha Rajesh Dikshit Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking [version 3; peer review: 2 approved] Gates Open Research |
author_facet |
Abhinendra Kumar Sharayu Mhatre Sheela Godbole Prabhat Jha Rajesh Dikshit |
author_sort |
Abhinendra Kumar |
title |
Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking [version 3; peer review: 2 approved] |
title_short |
Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking [version 3; peer review: 2 approved] |
title_full |
Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking [version 3; peer review: 2 approved] |
title_fullStr |
Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking [version 3; peer review: 2 approved] |
title_full_unstemmed |
Optimization of extraction of genomic DNA from archived dried blood spot (DBS): potential application in epidemiological research & bio banking [version 3; peer review: 2 approved] |
title_sort |
optimization of extraction of genomic dna from archived dried blood spot (dbs): potential application in epidemiological research & bio banking [version 3; peer review: 2 approved] |
publisher |
F1000 Research Ltd |
series |
Gates Open Research |
issn |
2572-4754 |
publishDate |
2019-11-01 |
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. |
url |
https://gatesopenresearch.org/articles/2-57/v3 |
work_keys_str_mv |
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