AN INNOVATIVE DNA EXTRACTION METHOD: WATER VERSUS COMMERCIAL BUFFERS

P. Mármol,B. Gómez,C. Gomes, C. Romero,C. Baeza-Richer, A. Hernández-Cordero, M. Martín-Arrebola, R. Rosell-Herrera,A.M. López-Parra,S. Palomo-Díez,E. Labajo-González,B Perea-Pérez,C. López-Matayoshi,E. Arroyo-Pardo

FORENSIC SCIENCE INTERNATIONAL GENETICS SUPPLEMENT SERIES(2019)

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Abstract
The main objective of DNA extraction is to obtain good quality genetic material in order to carry out its amplification, and corresponding analysis. Most laboratories tend to resort to commercial extraction buffers, which allow a simple and rigorous DNA extraction, with limited handling of the sample, but with high financial cost. As an alternative, we proposed to use water as reagent to extract DNA from blood spots, collected on Whaman (R) FTA (R) Cards (Sigma-Aldrich (R) ). We propose to take advantage of hypotonic solutions, in order to obtain a better efficiency in the extraction of genetic material (DNA). On this sense, 219 extractions were performed with Whatman (R) FTA (R) Cards (SigmaAldrich (R)) stained with blood, based on Garcia-Palacios a al., extraction method, with some modifications. Extractions were performed with two types of water (sterile (Lonza (TM)) and distilled (Milli-Q, MilliporeSigma (R)), as well as, with a commercial extraction kit (Prep-n-Go (TM), ThermoFisher (TM) Scientific, Foster City, USA). To evaluate and compare each reagent effectiveness, we amplified DNA extract with mitochondrial DNA Hypervariable regions I and II, and with nuclear DNA markers (autosomal STRs and X-InDels). Our preliminary results suggest that distilled water allows an extraction as effective, or even better, as that obtained with the commercial buffer. These preliminary results offer the possibility for laboratories to choose an inexpensive protocol for DNA extraction, avoiding expensive commercial buffers. Further research is needed to evaluate the performance of each method in blood samples, but also in other biological samples.
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Key words
DNA-extraction,Commercial buffer,Water,mtDNA,STRs
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