Determining SNP allele frequencies in DNA pools.
Single nucleotide polymorphisms (SNPs) are among the most common types of polymorphism used for genetic association studies. A method to allow the accurate quantitation of their allele frequencies from DNA pools would both increase throughput and decrease costs for large-scale genotyping. However, to date, most DNA pooling studies have concentrated on the use of microsatellite polymorphisms. In the case of SNPs that are restriction fragment length polymorphisms (RFLPs), studies have tended to use methods for the quantitation of allele frequency from pools that rely on densitometric evaluation of bands on an autoradiograph. Radiation-based methods have well-known drawbacks, and we present two alternative methods for the determination of SNP allele frequencies. For RFLPs, we used agarose gel electrophoresis of digested PCR products with ethidium bromide staining combined with densitometric analysis of gel images on a PC. For all types of SNP, we used allele-specific fluorescent probes in the Taqman assay to determine the relative frequencies of two different alleles. Both methods gave accurate and reproducible results, suggesting they are suitable for use in DNA pooling experiments.
Breen
G
G
University of Aberdeen, Scotland, UK. gbreen@abdn.ac.uk
Harold
D
D
Ralston
S
S
Shaw
D
D
St Clair
D
D
D000483:Alleles; D004247:DNA / Q000032:analysis*; D003720:Densitometry; D005456:Fluorescent Dyes; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length
D005456:Fluorescent Dyes; D004247:DNA