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 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.
Garcia-Sepulveda
Christian Alberto
CA
Laboratorio de Biología Molecular, Facultad de Medicina, Universidad Autónoma de San Luis Potosí, Avenida Venustiano Carranza #2405, Colonia Filtros las Lomas, 78210, San Luis Potosí, Mexico. christian.garcia@uaslp.mx
Carrillo-Acuña
Enrique
E
Guerra-Palomares
Sandra Elizabeth
SE
Barriga-Moreno
Montserrat
M
D001771:Blood Banks; D004247:DNA / Q000235:genetics / Q000302:isolation & purification*; D004587:Electrophoresis, Agar Gel; D005069:Evaluation Studies as Topic; D015894:Genome, Human / Q000235:genetics*; D006678:HIV / Q000235:genetics / Q000302:isolation & purification; D006515:Hepatitis B virus / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D007962:Leukocytes / Q000378:metabolism; D018895:Microsatellite Repeats / Q000235:genetics; D017720:Molecular Epidemiology / Q000379:methods*; D016133:Polymerase Chain Reaction; D013053:Spectrophotometry
D004247:DNA