Maxiprep genomic DNA extractions for molecular epidemiology studies and biorepositories
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Molecular epidemiology and genomic characterisation studies require the screening of large numbers of individuals to achieve statistical significance. Although many of the novel DNA extraction methods offer convenient, high-throughput capabilities, their use for the processing of larger sample volumes becomes very expensive. We are currently compiling the Mexican Genomic DNA Collection in order to address specific health priorities through molecular techniques. Our approach employs a low-cost laundry detergent based DNA extraction technique that maximizes DNA yield and quality. We have optimised four different modalities (maxiprep, midiprep, miniprep and microprep) for two different sources (leukocyte concentrates and whole blood). Our optimised protocol produces 4.5 mg of DNA from 15 ml of blood-bank discarded leukocyte concentrates with spectrophotometric quality, genomic integrity and PCR suitability that rivals that of phenol–chloroform extracted samples. We present evidence of many PCR applications that we have carried out on samples extracted with this technique including Killer-cell Immunoglobulin-like Receptor genotyping, Short Tandem Repeat profiling as well as nucleic acid screening for hepatitis B and human immunodeficiency type-1 viruses. This paper highlights many of the advantages that this DNA extraction technique provides over existing methodologies, whether it is used to establish large genomic DNA collections (as was our main intention) or as a routine DNA extraction method for PCR applications.
KeywordsMolecular epidemiology DNA banking DNA extraction Methods Biorepository
The authors wish to thank Dr. Flavio J. Martínez Morales of the Pharmacology Laboratory for his valuable support and for providing equipment without which this study would not have been possible. To Dr. Daniel E. Noyola Cherpitel for proofreading this paper and to Hugo Contreras Treviño for helping with the phenol–chloroform extractions. This work was supported by funds from the Mexican Consejo Nacional de Ciencia y Tecnología (CONACYT) grant number #55360 and the Programa de Mejoramiento del Profesorado (PROMEP) of the Universidad Autónoma de San Luis Potosi (UASLP-PTC-155).
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