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

Gates Open Research

article

Abhinendra Kumar¹  Sharayu Mhatre¹  Sheela Godbole³  Prabhat Jha⁴  Rajesh Dikshit¹ 

[1] Centre for Cancer Epidemiology, Tata Memorial Centre;Homi Bhabha National Institute, Training School Complex;Department of Biostatistics and Epidemiology, National AIDS Research Institute;Li Ka Shing Knowledge Institiute, St Michael's Hospital, Center for Global Health Research

关键词: Dried blood spot (DBS);    Whatman 903 cards;    FTA cards;    Human genomic DNA;    Bio-banking;    Epidemiology;   

DOI  :  10.12688/gatesopenres.12855.3

学科分类：电子与电气工程

来源: American Journal Of Pharmtech Research

【 摘 要 】

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.

【 授权许可】

CC BY