Table 1 Primer details for quantitative RT-PCR. EEF1A, Elongation factor 1α; COL2A1, collagen type II alpha 1; COL10A1, collagen type X alpha 1; ALP, alkaline phosphatase; SOX9, sex-determining region Y-box 9
Gene | primer sequences (5'-3') | annealing temperature (°C) | product size (base pairs) | cycles | MgCl2 |
|---|---|---|---|---|---|
Housekeeping gene for internal control | |||||
EEF1A | Sense: AGGTGATTATCCTGAACCATCC Antisense: AAAGGTGGATAGTCTGAGAAGC | 54.0 | 234 | 25 | 1 x |
Chondrogenic marker genes | |||||
COL2A1 | Sense: TTTCCCAGGTCAAGATGGTC Antisense: CTTCAGCACCTGTCCACCA | 58.0 | 374 | 35 | 1x |
SOX9 | Sense: ATCTGAAGAAGGAGAGCGAG Antisense: TCAGAAGTCTCCAGAGCTTG | 58.0 | 263 | 35 | 1x |
Chondrogenic hypertrophy marker genes | |||||
COL10A1 | Sense: CCCTTTTTGCTGCTAGTATCC Antisense: CTGTTGTCCAGGTTTTCCTGGCAC | 54.0 | 468 | 25 | 1x |
ALP | Sense: TGGAGCTTCAGAAGCTCAACACCA Antisense: ATCTCGTTGTCTGAGTACCAGTCC | 51.0 | 454 | 35 | 1x |