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Function of reference genes, primer sequences, optimal primer concentrations and reaction efficiencies of qPCR experiments. |
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| Symbol |
Function |
Primer sequence |
nM |
Efficiency |
|
|
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| ACTB |
Cytoskeletal structural protein |
fw: CTGGAACGGTGAAGGTGACA rv: AAGGGACTTCCTGTAACAATGCA |
300 100 |
1.94 ± 0.01 |
| GAPDH |
Oxidoreductase in glycolysis and gluconeogenesis |
fw: GGAGTCCACTGGCGTCTTCAC rv: GAGGCATTGCTGATGATCTTGAGG |
300 600 |
1.94 ± 0.01 |
| B2M |
Beta-chain of major histocompatibility complex class I molecules |
fw: TGCTGTCTCCATGTTTGATGTATCT rv: TCTCTGCTCCCCACCTCTAAGT |
300 900 |
2.03 ± 0.04 |
| HPRT1 |
Purine synthesis in salvage pathway |
fw: TGACACTGGCAAAACAATGCA rv: GGTCCTTTTCACCAGCAAGCT |
300 900 |
1.94 ± 0.03 |
| SDHA |
Electron transporter in the TCA cycle and respiratory chain |
fw: TGGGAACAAGAGGGCATCTG rv: CCACCACTGCATCAAATTCATG |
300 900 |
2.00 ± 0.01 |
| YWHAZ |
Signal transduction by binding to phosphorylate serine residues |
fw: ACTTTTGGTACATTGTGGCTTCAA rv: CCGCCAGGACAAACCAGTAT |
600 600 |
1.98 ± 0.01 |
|
Optimal primer concentrations were determined using a matrix of forward and reverse primers varying in concentration from 50 nM to 900 nM. Combinations that gave the lowest Ct value and the highest ΔRn value were selected. qPCR efficiencies for each primer pair were derived from standard curves (n = 3) using five-fold dilution series covering a 4 to 5 log dynamic range starting from one randomly selected undiluted cDNA sample of the untreated control group. | ||||
Toegel et al. BMC Molecular Biology 2007 8:13 doi:10.1186/1471-2199-8-13 |
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