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Apolipoprotein E genotyping method by Real Time PCR, a fast and cost-effective alternative to the TaqMan® and FRET assays
October 2009
October 2009
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Abstract
The apolipoprotein E gene (APOE) polymorphism genotyping has an allegedly important
predictive value for coronary heart disorders and Alzheimer's disease. We developed
a simple, fast, cost-effective and suited for high-throughput protocol for determining
APOE genotypes by Real Time PCR monitored by SYBR Green. The method is based on differential
amplification by allele-specific primers. These primers have variations in their 3'-end
nucleotides such that are specific for one of the two variants in each polymorphic
position. By this protocol, we obtained a 100% concordance with the APOE genotypes
determined by sequencing analysis. The main advantages of this method are its relative
simplicity and the reduced cost compared to other methodologies, such as the TaqMan
and FRET assays.