Figure 1

Validating RT-PCR amplification across the CAG repeat in HD patient-derived fibroblasts. Wild-type and mutant HTT were separated by gel electrophoresis. (A) Standard curve of wild-type and mutant HTT RT-PCR products with increasing PCR cycles from gDNA derived from post-mortem brain tissue of 7 HD patients. PCR linearity was evaluated by determining the individual linear regression coefficients (r ²) of the band intensities of wild-type and mutant HTT expression versus the number of PCR cycles, n = 7. (B) PCR products from cDNA of 4 HD (GM00305, GM02173, GM04022, GM04855) fibroblasts. CAG repeat sizes for the wild-type (lower band) and mutant alleles (upper band) are indicated below each lane. gDNA was used to examine differences in PCR amplification between the wild-type and mutant product due to the CAG repeat expansion. (C) RT-PCR products with input: cDNA (+RT), cDNA lacking reverse transcriptase (−RT) and gDNA of one control (GM04204). (D) Whisker boxplot of RT-PCR from HD patient-derived fibroblasts, comparing wild-type and mutant HTT mRNA expression levels, relative to gDNA. Line = mean, pair wise differences were evaluated using linear mixed model, n = 4.