Gene | RT-PCR primer sequences (5'-3') | Annealing temp. (°C) | Product size (bp) | Cycles |
---|---|---|---|---|
Chondrogenic markers | ||||
COL II | Sense: TTTCCCAGGTCAAGATGGTC Antisense: CTTCAGCACCTGTC CACCA | 58 | 374 | 35 |
AGC | Sense: TGAGGAGGGCTGGAACAAGTACC Antisense: GGAGGTGGTAATTGCAGGGAACA | 54 | 392 | 30 |
COMP | Sense: CAGGACGACTTTGATGCAGA Antisense: AAGCTGGAGCTGTCTGGTA | 54 | 312 | 32 |
FMD | Sense: CTTACCCCTATGGGGTGGAT Antisense: GTACATGGCCGTGAGGAAGT | 54 | 389 | 35 |
SOX9 | Sense: ATCTGAAGAAGGAGAGCGAG Antisense: TCAGAAGTCTCCAGAGCTTG | 58 | 263 | 35 |
SOX9 (rt) | Sense: GGA GTGGAAGTTACTGACTGATG Antisense: AGGCGTTTTGCTTCGTCAATG | 55 | 60 | Â |
Hypertrophy and osteogenic markers | ||||
COL I | Sense: GGACACAATGGATTGCAAGG Antisense: TAACCACTGCTCCACTCTGG | 54 | 461 | 32 |
COL X | Sense: CCCTTTTTGCTGCTAGTATCC Antisense: CTGTTGTCCAGGTTTTCCTGGCAC | 54 | 468 | 25 |
OP | Sense: ACGCCGACCAAGGAAAACTC Antisense: GTCCATAAACCACACTATCACCTCG | 51 | 483 | 35 |
ALP (rt) | Sense: TGGAGCTTCAGAAGCTCAACACCA Antisense: ATCTCGTTGTCTGAGTACCAGTCC | 51 | 454 | Â |
IHH | Sense: GAGGAGTCCCTGCATTATGA Antisense: CAGGAAAATGAGCACATCGC | 54 | 321 | 30 |
RUNX2 | Sense: ACAGATGATGACACTGCCACC Antisense: CATAGTAGAGATATGGAGTGCTGC | 55 | 324 | 35 |
Internal control | ||||
EF1α | Sense: AGGTGATTATCCTGAACCATCC Antisense: AAAGGTGGATAGTCTGAGAAGC | 54 | 234 | 25 |