cDNA samples from human brain region tissues and genomic samples, both from individuals who were heterozygous for the PPP3CA 5′UTR tri-nucleotide repeat (GCC), were used for allele-specific expression assays. Genomic DNA and cDNA samples from medial temporal gyrus (n = 13) and/or medial frontal gyrus (n = 14) were added to 384-well plates containing 1× reaction buffer (Stratagene), 0.8 mM dNTPs (Stratagene), CYBR-1 nucleic acid green stain dye (Biowhittaker Molecular Applications), 1× Rox reference dye (Invit-rogen), 5% DMSO, 20 uM each of PPP3CA_EX1F4 and PPP3CA_EX1R5, and Herculase enzyme (Stratagene). Reactions used an ABI Prism 7900HT Sequence Detection System with 95°C denaturation for 10 min followed by 45 cycles of 94°C for 0.5 min, 62°C for 0.5 min, 72°C for 0.75 min, and then terminal 10 min 72°C incubation. Dissociation, involved a hold at 50°C for 5 min, followed by one cycle of 95°C for 0.25 min, 60°C for 0.25 min, and 95°C for 0.25 min, and was used to degrade short fragment primer dimers. The linear range for each sample was identified using SDS v2.1 software. Using the determined linear
