title: Analysis of clinically relevant single-nucleotide polymorphisms by use of microelectronic array technology.
abstract: BACKGROUND
  Microelectronic DNA chip devices represent an emerging technology for genotyping. We developed methods for detection of single-nucleotide polymorphisms (SNPs) in clinically relevant genes.
  
  
  METHODS
  Primer pairs, with one containing a 5'-biotin group, were used to PCR-amplify the region encompassing the SNP to be interrogated. After denaturation, the biotinylated strand was electronically targeted to discrete sites on streptavidin-coated gel pads surfaces by use of a Nanogen Molecular Workstation. Allele-specific dye-labeled oligonucleotide reporters were used for detection of wild-type and variant sequences. Methods were developed for SNPs in genes, including factor VII, beta-globin, and the RET protooncogene. We genotyped 331 samples for five DNA variations in the factor VII gene, >600 samples from patients with beta-thalassemia, and 15 samples for mutations within the RET protooncogene. All samples were previously typed by various methods, including DNA sequence analysis, allele-specific PCR, and/or restriction enzyme digestion of PCR products.
  
  
  RESULTS
  Analysis of amplified DNA required 4-6 h. After mismatched DNA was removed, signal-to-noise ratios were >5. More than 940 samples were typed with the microelectronic array platform, and results were totally concordant with results obtained previously by other genotyping methods.
  
  
  CONCLUSIONS
  The described protocols detect SNPs of clinical interest with results comparable to those of other genotyping methods.
authors.affiliation: Department of Pediatrics, The Children's Hospital of Philadelphia, University of Pennsylvania School of Medicine, Philadelphia, PA 19104, USA.
authors.forename: Rosa
authors.initials: R
authors.lastname: Santacroce
authors.forename: Antonia
authors.initials: A
authors.lastname: Ratti
authors.forename: Francesco
authors.initials: F
authors.lastname: Caroli
authors.forename: Barbara
authors.initials: B
authors.lastname: Foglieni
authors.forename: Alessandro
authors.initials: A
authors.lastname: Ferraris
authors.forename: Laura
authors.initials: L
authors.lastname: Cremonesi
authors.forename: Maurizio
authors.initials: M
authors.lastname: Margaglione
authors.forename: Marco
authors.initials: M
authors.lastname: Seri
authors.forename: Roberto
authors.initials: R
authors.lastname: Ravazzolo
authors.forename: Gabriella
authors.initials: G
authors.lastname: Restagno
authors.forename: Bruno
authors.initials: B
authors.lastname: Dallapiccola
authors.forename: Eric
authors.initials: E
authors.lastname: Rappaport
authors.forename: Eleanor S
authors.initials: ES
authors.lastname: Pollak
authors.forename: Saul
authors.initials: S
authors.lastname: Surrey
authors.forename: Maurizio
authors.initials: M
authors.lastname: Ferrari
authors.forename: Paolo
authors.initials: P
authors.lastname: Fortina
mesh_terms: D005167:Factor VII / Q000235:genetics; D005914:Globins / Q000235:genetics; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D015513:Oncogene Proteins / Q000235:genetics; D020641:Polymorphism, Single Nucleotide; D051096:Proto-Oncogene Proteins c-ret; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics
chemical_list: D015513:Oncogene Proteins; D005167:Factor VII; D005914:Globins; D051096:Proto-Oncogene Proteins c-ret; C099282:RET protein, human; D020794:Receptor Protein-Tyrosine Kinases