title: Quantitative approach to single-nucleotide polymorphism analysis using MALDI-TOF mass spectrometry.
abstract: Pooling of DNA samples before genotyping is a valuable means of streamlining large-scale genotyping efforts in disease association studies, single-nucleotide polymorphism (SNP) validation or mutant allele screening programs. In this report, we explore the application of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to quantitative analysis of SNPs. The measurements are based on MALDI-TOF MS analysis of primer extension assays performed on standard mixtures of pooled PCR products at several test loci. The inherent high molecular weight resolution of MALDI-TOF MS conveys high specificity and good signal-to-noise ratio for performing accurate quantitation. The methods described maximize the sensitivity and quantitative capacity of MALDI-TOF MS while preserving the throughput and economic advantages of the MALDI-TOF platform. Using the format described, we demonstrate that allele frequencies as low as 5% can be detected quantitatively and unambiguously.
authors.affiliation: Applied Biosytems, Framingham, MA, USA. philip_ross@pbio.com
authors.forename: P
authors.initials: P
authors.lastname: Ross
authors.forename: L
authors.initials: L
authors.lastname: Hall
authors.forename: L A
authors.initials: LA
authors.lastname: Haff
mesh_terms: D000483:Alleles; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics; D004247:DNA / Q000032:analysis; D005634:Fructose-Bisphosphate Aldolase / Q000235:genetics; D005787:Gene Frequency; D005838:Genotype; D016133:Polymerase Chain Reaction; D020641:Polymorphism, Single Nucleotide / Q000235:genetics; D011973:Receptors, LDL / Q000235:genetics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D011973:Receptors, LDL; D004247:DNA; D003577:Cytochrome P-450 Enzyme System; D005634:Fructose-Bisphosphate Aldolase