Table 2.
Primer sequences for target and reference genes used in RT–qPCR assays
| Gene | Accession genea | Sequence 5′–3′ | Product size (bp) | Source |
|---|---|---|---|---|
| GAPDH | NM_001034034* | GGCGTGAACCACGAGAAGTATAA CCCTCCACGATGCCAAAGT |
119 | [25] |
| HPRT1 | AF176419* | TGCTGAGGATTTGGAGAAGG CAACAGGTCGGCAAAGAACT |
154 | [26] |
| 18S | NA* | NA | NA | PD |
| COL2A1 | NM_001113224* | TGGCTGACCTGACCTGAC GGGCGTTTGACTCACTCC |
187 | BD |
| ACAN | NM_173981* | CCAACGAAACCTATGACGTGTACT GCACTCGTTGGCTGCCTC |
107 | [27] |
| T | NM_001192985 | AAGAACGGCAGGAGGATGT CCAGCAGGAAGGAGTACATG |
82 | PD |
| KRT8 | NM_001159615* | TGCTGGAGGGAGAGGAGAG CTGTAGTTGAAGCCTGGAGTC |
118 | PD |
| KRT18 | NM_001192095* | AAGAACGGCAGGAGGATGT CCAGCAGGAAGGAGTACATG |
82 | PD |
| KRT19 | XM_003131437 | AACCAGGAGTACCAGCATCT CTTGATGATGGTCAGGTTGTTG |
117 | PD |
GAPDH glyceraldehyde-3-phosphate dehydrogenase, HPRT1 hypoxanthine phosphoribosyltransferase, COL2A1 collagen, type II, alpha 1, ACAN aggrecan, T Brachyury, KRT8 cytokeratin 8, KRT18 cytokeratin 18, KRT19 cytokeratin 19, NA Information not provided by the manufacturer, BD Primers designed with Beacon designer software (Premier Biosoft, Palo Alto, CA, USA) and ordered from Sigma, PD primers designed by and ordered from PrimerDesign Ltd (Southampton, UK); other primers ordered from Sigma
* Primers tested and validated for both bovine and porcine gene
aThe accession number is the GenBank™ accession number