- Rapid DNA Extraction from Dried Blood Spots on Filter Paper: Potential Applications in Biobanking
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Eun-Hye Choi, Sang Kwang Lee, Chunhwa Ihm, Young-Hak Sohn
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Osong Public Health Res Perspect. 2014;5(6):351-357. Published online December 31, 2014
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DOI: https://doi.org/10.1016/j.phrp.2014.09.005
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Abstract
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- Objectives
Dried blood spot (DBS) technology is a microsampling alternative to traditional plasma or serum sampling for pharmaco- or toxicokinetic evaluation. DBS technology has been applied to diagnostic screening in drug discovery, nonclinical, and clinical settings. We have developed an improved elution protocol involving boiling of blood spots dried on Whatman filter paper. Methods
The purpose of this study was to compare the quality, purity, and quantity of DNA isolated from frozen blood samples and DBSs. We optimized a method for extraction and estimation of DNA from blood spots dried on filter paper (3-mm FTA card). A single DBS containing 40 μL blood was used. Results
DNA was efficiently extracted in phosphate-buffered saline (PBS) or Tris-EDTA (TE) buffer by incubation at 37°C overnight. DNA was stable in DBSs that were stored at room temperature or frozen. The housekeeping genes GAPDH and beta-actin were used as positive standards for polymerase chain reaction (PCR) validation of general diagnostic screening. Conclusion
Our simple and convenient DBS storage and extraction methods are suitable for diagnostic screening by using very small volumes of blood collected on filter paper, and can be used in biobanks for blood sample storage.
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