Sequence detector software sds version 1.6

Gene expression was measured using commercial TaqMan assays (Applied Biosystems; Table 2). The expression of GAD67 and selected GABA-A receptor subunits (Table 2) were measured in preamplified cDNA derived from the gray matter of the ACC from each subject. In addition the expression levels of the housekeeping genes glyceraldehyde-3-phosphate dehydrogenase (GAPDH), cyclophilin (PPIA), and glucuronidase beta (GUSB) were measured. Assays for each target gene were performed in duplicate in 96-well optical plates using a Stratagene MX3000P instrument (Stratagene, La Jolla, California) and Sequence Detector Software (SDS version 1.6; PE Applied Biosystems). The relative standard curve method was used for these analyses as described previously (Sodhi et al., 2011 (link)). Briefly, standard curves were generated for each target assay and for each endogenous control assay using a calibration curve and the geometric mean of GAPDH, PPIA, and GUSB expression was used for normalization of the target genes according to Applied Biosystems instructions (Guide to Performing Relative Quantitation of Gene Expression Using Realtime Quantitative PCR, Applied Biosystems).