Table 1.
Primers and antibodies used in this study.
| Primer Names | Sequences (5´ to 3´) | Purpose | Cycling Conditions |
| PCR | Genomic DNA amplification data | 95 °C for 5 min, 35 cycles of 95 °C 30 s, 60 °C 30 s and 72 °C 1 to 4 min | |
| intron 1F | cagtctagacagcagggcaa | ||
| exon 2R | agtaggatgtgccccaggag | ||
| exon 1F2 | cggcctctcagttgggaaa | ||
| exon 2R2 | ggttgccaaggacacga | ||
| exon 2F | tgtaggagctgtccaggcc | ||
| exon 5R2 | ggcattgctgtttagcag | ||
| intron 4F | gatctcggtcttgaaagc | ||
| exon 10R2 | actcagccaccatgtcctcc | ||
| exon 10F | actgccttccccgacttca | ||
| exon 15R | tggaacagtgtgtagaggtg | ||
| exon 15F | cgtacagcttcagcgag | ||
| exon 16R | tgcaggtcgtaccatgtg | ||
| exon 16F | caaggactctagcacctgg | ||
| exon 20R | gaatctcccaagtcctgtgadata | ||
| RT-PCR | GAA transcript amplification | 95 °C for 5 min 35 cycles of 95 °C 30 s, 60 °C 30 s and 72 °C 1 min | |
| exon 1F | ggaaactgaggcacggagcg | ||
| exon 5R | ggaccacatccatggcattgc | ||
| exon 4F | gtatatcacaggcctcgccg | ||
| exon 10R | ctggtcatggaactcagcca | ||
| exon 9F | gggggttttcatcaccaacga | ||
| exon 17R | ctgccaagggcctctactgg | ||
| exon 16F | caaggactctagcacctgg | ||
| exon 20R | gaatctcccaagtcctgtga | ||
| RT-qPCR | Full-length GAA transcript (exon 1-2), amplification | 95 °C for 1 min, 40 cycles of 95 °C 3 s, 60 °C 15 s and 72 °C 30 s | |
| exon 1F(q) | tgggaaagctgaggttgtcg | ||
| exon 1-2R(q) | tcctacaggcccgctcc | ||
| TBP transcript amplification | |||
| exon 1-2F(q) | tctttgcagtgacccagcatcac | ||
| exon 2R(q) | cctagagcatctccagcacactct | ||
| Antibodies | Catalogue no. | Source | Dilution |
| rabbit anti-GAA | 137068 | Abcam, Melbourne, Australia | 1:1000 |
| mouse monoclonal anti-β-tubulin | E7 | DSHB, Iowa City, Iowa | 1: 5000 |
| mouse monoclonal anti-β-actin | A5316 | Sigma-Aldrich, Castle Hill NSW | 1: 500,000 |