title: Validation of a fast and low-cost alkaline lysis method for gDNA extraction in a pharmacogenetic context.
abstract: OBJECTIVE
  Translation of pharmacogenetic findings from the research laboratory to the clinical practice demands simple and efficient procedures. In this sense, we evaluated the suitability of a modified protocol for genomic DNA extraction based on alkaline lysis of cells.
  
  
  METHODS
  Dried blood samples were obtained from 48 patients diagnosed with colorectal cancer. A total of 11 mutations in the dihydropyrimidine dehydrogenase gene and related to 5-fluorouracil toxicity were searched by amplicon sequencing and real-time PCR with fluorescent probes.
  
  
  RESULTS
  Genomic DNA extracted with the alkaline lysis method, both from dried blood samples and buccal swabs, fulfilled the quality requirements of the two genotyping methods assayed, which yielded 100 % concordant results for 11 genetic variants with relevance to cancer chemotherapy.
  
  
  CONCLUSIONS
  The assessed protocol has shown to be a very fast and economical approach to perform genetic analyses in the clinical laboratory for pharmacological purposes.
authors: Pharmacy Department, University Hospital of the Canary Islands (HUC), Tenerife, Spain.
authors: Ruth
authors: R
authors: Ramos-Díaz
authors: Fernando
authors: F
authors: Gutiérrez-Nicolás
authors: Gloria J
authors: GJ
authors: Nazco-Casariego
authors: Itamar
authors: I
authors: González-Perera
authors: José A
authors: JA
authors: Pérez-Pérez
mesh_terms: D000468:Alkalies / Q000737:chemistry; D015179:Colorectal Neoplasms / Q000097:blood* / Q000188:drug therapy / Q000235:genetics*; D004273:DNA, Neoplasm / Q000097:blood / Q000235:genetics / Q000302:isolation & purification*; D042943:Dihydrouracil Dehydrogenase (NADP) / Q000235:genetics; D005472:Fluorouracil / Q000633:toxicity; D060005:Genotyping Techniques; D006801:Humans; D009154:Mutation; D010597:Pharmacogenetics; D011446:Prospective Studies; D015203:Reproducibility of Results
chemical_list: D000468:Alkalies; D004273:DNA, Neoplasm; D042943:Dihydrouracil Dehydrogenase (NADP); D005472:Fluorouracil