For SNP rs1824024, the SNP showing statistical significance for its association with the P300 trajectories in the present analysis, a 1 pmol of each of the unmodified forward primer 5′-GTGGGCCTCAGAGAGACCATA-3′ and the biotinylated reverse primer 5′-GGCAGTAGGAAGTAAATGGATCA-3′. (primers for the other six SNPs are available on request) was used. Thermal cycling included 45 cycles at an annealing temperature of 60 degrees. The Biotage workstation was used to isolate the biotinylated single strand from the double strand PCR products. The isolated product was then sequenced using the complimentary sequencing primer 5′-AGACCATACCGAGGG-3′. At the polymorphic site, the minor allele was detected by the presence of a G nucleotide whereas the major allele was detected by the presence of a T nucleotide.
