Table 2.
Primer details for semiquantitative RT-PCR
| Gene | Primer sequences (5′-3′) | Annealing temperature (°c) | Product size (base pairs) | Cycles | MgCl2 |
|---|---|---|---|---|---|
| Housekeeping gene for internal control | |||||
| EEF1α | Sense: AGGTGATTATCCTGAACCATCC Antisense: AAAGGTGGATAGTCTGAGAAGC | 54.0 | 234 | 21 | 1 x |
| Adipogenic marker genes | |||||
| LPL |
Sense: GAGATTTCTCTGTATGGCACC Antisense: CTGCAAATGAGACACTTTCTC |
51.0 | 239 | 30 | 1x |
| PPARγ |
Sense: GCTGTTATGGGTGAAACTCTG Antisense: ATAAGGTGGAGATGCAGGCTC |
61.0 | 297 | 35 | 1x |
| Osteogenic marker genes | |||||
| Col I |
Sense: GGACACAATGGATTGCAAGG Antisense: TAACCACTGCTCCACTCTGG |
55.0 | 461 | 22 | 2x |
| Col X |
Sense: CCCTTTTTGCTGCTAGTATCC Antisense: CTGTTGTCCAGGTTTTCCTGGCAC |
54.0 | 468 | 40 | 1x |
| ALP |
Sense: TGGAGCTTCAGAAGCTCAACACCA Antisense: ATCTCGTTGTCTGAGTACCAGTCC |
51.0 | 454 | 33 | 1x |
| OC |
Sense: ATGAGAGCCCTCACACTCCTC Antisense: GCCGTAGAAGCGCCGATAGGC |
62.0 | 293 | 35 | 2x |
| Chondrogenic marker genes | |||||
| AGG |
Sense: GCCTTGAGCAGTTCACCTTCAntisense: CTCTTCTACGGGGACAGCAG |
54.0 | 400 | 35 | 1x |
| Col II |
Sense: TTTCCCAGGTCAAGATGGTC Antisense: CTTCAGCACCTGTCCACCA |
51.0 | 155 | 31 | 1x |
| Sox-9 |
Sense: ATCTGAAGAAGGAGAGCGAG Antisense: TCAGAAGTCTCCAGAGCTTG |
60.0 | 263 | 31 | 1x |
EEF 1α Elongation factor 1α, LPL Lipoproteinlipase, PPARγ Peroxisome proliferator-activated receptor γ, Col I Collagen type I, Col X Collagen type X, ALP Alkaline phosphatase, OC Osteocalcine, AGG Aggrecane, Col II Collagen type II, Sox-9 Sex-determining region Y-box 9