Table 1 |
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|
Primer sequences and product sizes, for semi-quantitative and real-time RT-PCR |
||||
|
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 |
|
|
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 |
|
|
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 |
|
|
||||
|
rt: primer pairs, that have been used for real-time PCR only. AGC = aggrecan core protein; ALP = alkaline phosphatase; COL = collagen; COMP = cartilage oligomeric matrix protein; EF1α = elongation factor 1α; FMD = fibromodulin; IHH = indian hedgehog; OP = osteopontin; RUNX2 = runt-related transcription factor 2; SOX9 = SRY (sex determining region Y) - box9. |
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|
Steinert et al. Arthritis Research & Therapy 2009 11:R148 doi:10.1186/ar2822 |
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