Table 3 Oligonucleotide sequences, modifications, final concentrations, and assay-specific annealing temperatures
Gene | Forward Primer (5′ → 3 ′) | Reverse Primer (5′ → 3 ′) | Probe (5′ → 3 ′)¥ | Annealing temperature |
|---|---|---|---|---|
CTLA4 | 0.4 µM CTCATGTACCCACCGCCATACT | 0.4 µM TTGATGGGAATAAAATAAGGCTGAA | 0.2 µM 6-FAM-CAGATTTATGTAATTGATCCAGAACCGTGCC-BHQ-1 | 62 °C |
ACTB | 0.2 µM ATGTGGCCGAGGACTTTGATT | 0.2 µM AGTGGGGTGGCTTTTAGGATG | 0.16 µM HEX-GAAATRMGTKGTTACAGGAAGTCCCT-BHQ-1 | 58 °C |
ALAS1 | 0.2 µM TAATGACTACCTAGGAATGAGTCG | 0.2 µM CCATGTTGTTTCAAAGTGTCCA | 0.16 µM 6-FAM-TAACTGCCCCACACACCCGT-BHQ-1 | 62 °C |
GAPDH | 0.2 µM TGCACCACCAACTGCTTAGC | 0.2 µM GGCATGGACTGTGGTCATGAG | 0.16 µM 6-FAM-CTGGCCAAGGTCATCCATGACAACT-BHQ-1 | 58 °C |
SDHA | 0.2 µM TCGCTCTTGGACCTGGT | 0.2 µM TGGAGGGACTTTATCTCCAG | 0.16 µM 6-FAM-ATCGAAGAGTCATGCAGGCC-BHQ-1 | 62 °C |
HPRT1 | 0.2 µM TGACACTGGCAAAACAATGCA | 0.2 µM GGTCCTTTTCACCAGCAAGCT | 0.16 µM 6-FAM-TGCTTTCCTTGGTCAGGCAGTAT-BHQ-1 | 62 °C |