Biorepository; DNA banking; DNA extraction; Methods; Molecular epidemiology chloroform; genomic DNA; killer cell immunoglobulin like receptor; nucleic acid; phenol; DNA; microsatellite DNA; article; bioinformatics; controlled study; DNA extraction; genotype; health care planning; Hepatitis B virus; human; human cell; Human immunodeficiency virus 1; leukocyte; molecular biology; molecular epidemiology; polymerase chain reaction; separation technique; short tandem repeat; spectrophotometry; agar gel electrophoresis; blood bank; evaluation; genetics; human genome; Human immunodeficiency virus; isolation and purification; metabolism; methodology; Blood Banks; DNA; Electrophoresis, Agar Gel; Evaluation Studies as Topic; Genome, Human; Hepatitis B virus; HIV; Humans; Leukocytes; Microsatellite Repeats; Molecular Epidemiology; Polymerase Chain Reaction; Spectrophotometry
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 bloodbank 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. © Springer Science%2bBusiness Media B.V. 2009.
Maxiprep genomic DNA extractions for molecular epidemiology studies and biorepositories
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 bloodbank 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. © Springer Science Business Media B.V. 2009.
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Consejo Nacional de Ciencia y Tecnología, CONACYT: 55360
Universidad Autónoma de San Luis Potosí, UASLP: UASLP-PTC-155
Generically Dependent Continuant
Fondo Nacional de Ciencia y Tecnología, FONACYT
RDF description of Maxiprep genomic DNA extractions for molecular epidemiology studies and biorepositories - http://orbis.uaslp.mx/vivo/individual/scopus2473
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