Table 2 Primer list and sequences of 12 candidate reference genes for real-time PCR
Gene name | Gene product | Function | Accession number | Primer sequence | Primer efficiency |
|---|---|---|---|---|---|
GAPDH (98 bp) | Glyceraldehyde 3-phosphate dehydrogenase | Enzyme of glycolysis | NM_001190390.1 | F:5’ ACAGTCAAGGCAGAGAACGG 3′ R:5’ CCAGCATCACCCCACTTGAT 3’ | 97.0% |
ALAS1 (123 bp) | Delta-aminolevulinate synthase 1 | Enzyme of heme biosynthetic pathway | XM_004018407.3 | F:5’ CACTGCCCCAGTCACATCAT 3′ R:5’ GGGCACTGTGGGGTAATTGA 3’ | 95.5% |
HPRT (102 bp) | Hypoxanthine-guanine phosphoribosyl-transferase | Enzyme of purine salvage pathway | XM_015105023.1 | F:5’ TTCTTTGCCGACCTGTTGGA 3′ R:5’ TCACCTGTTGACTGGTCGTT 3’ | 92.5% |
EF-2 (106 bp) | Elongation factor 2 | required for the translocation step in protein synthesis | XM_012178077.2 | F:5’ GTTGTGAAGGCCTACCTCCC 3′ R:5’ GCCAGTGGTCAAACACACAC 3’ | 99.5% |
G6PDH (106 bp) | Glucose-6-phosphate dehydrogenase | Enzyme of pentose phosphate pathway | NM_001093780.1 | F:5’ ATTGTGGAGAAGCCCTTCGG 3′ R:5’ GGTAGTGGTCGATGCGGTAG 3’ | 94.5% |
ACTB (95 bp) | Beta-actin | Cytoskeletal structural protein | NM_001009784.2 | F:5’ GCAGATGTGGATCAGCAAGC 3′ R:5’ GGGTGTAACGCAGCTAACAG 3’ | 92.0% |
RPL19 (126 bp) | Ribosomal protein L19 | Ribosomal protein | XM_004012836.2 | F:5’ AGCCTGTGACTGTCCATTCC 3′ R:5’ ACGTTACCTTCTCGGGCATT 3’ | 99.0% |
B2M (91 bp) | Beta-2 microglobulin | Beta-chain of MHC-I | NM_001009284.2 | F:5’ CCTTGGTCCTTCTCGGGCTG 3′ R:5’ TCTGGCGGGTGTCTTGAGTAT 3’ | 98.5% |
YWHAZ (124 bp) | Tyrosine 3-monooxygenase/ tryptophan 5-monooxygenase activation protein, zeta | Signal transduction | NM_001267887.1 | F:5’ GATGAAGCCATTGCTGAACTTGA 3′ R:5’ CAGCTTCGTCTCCTTGGGTA 3’ | 97.0% |
SDHA (104 bp) | Succinate dehydrogenase | Enzyme of mitochondrial respiratory chain | XM_012097183.1 | F:5’ GAGTTCGTGCAGTTCCACCC 3′ R:5’ CTCTCACCCTGGCTGTTGAT 3’ | 91.5% |
PGK1 (104 bp) | Phosphoglycerate kinase 1 | Enzyme of glycolysis | NM_001142516.1 | F:5’ CCTCTGGCATACCTGTTGGC 3′ R:5’ CACCCACAGGTCCATTCCAC 3’ | 92.5% |
TFRC (96 bp) | Transferrin receptor | Transmembrane glycoprotein | XM_004003001.2 | F:5’ ACCTCAAATCAGCGCTGTCA 3′ R:5’ CAGCCTCACGTGGGACATAA 3’ | 93.5% |