Table 1 Primer, sequences, and protocols for quantitative real-time polymerase chain reaction
From: Fibroblast-like cells change gene expression of bone remodelling markers in transwell cultures
Primer | Sequenzen (5′-3′) | Primer concentration (nM) | Amplification | Efficiency (E = 10−1/slope) |
|---|---|---|---|---|
ALP | TCAAGGGTCAGCTCCACCACA ATTGGCCTTCACCCCACACA | 300 | 95 °C 10 s, 60 °C 10 s, 72 °C 30 s for 45 cycles | 2.01 |
Collagen 1A1 | GCTTCCCTGGTCTTCCTG TCTCACCACGGTCACCCT | 500 | 95 °C 10 s, 65 °C 10 s, 72 °C 15 s for 40 cycles | 2.01 |
GAPDH | TGCACCACCAACTGCTTAGC GGCATGGACTGTGGTCATGAG | 300 | 95 °C 10 s, 60 °C 10 s, 72 °C 15 s for 40 cycles | 2.04 |
Cathepsin K | TTCCCGCAGTAATGACACC TTTCCCCAGTTTTCTCCCC | 500 | 95 °C 10 s, 63 °C 10 s, 72 °C 20 s for 40 cycles | 1.83 |
M-CSF | CCGAGGAGGTGTCGGAGTAC AATTTGGCACGAGGTCTCCAT | 300 | 95 °C 10 s, 60 °C 10 s, 72 °C 15 s for 40 cycles | 2.01 |
MMP 13 | GACTTCACGATGGCATTGCTG GCATCAACCTGCTGAGGATGC | 300 | 95 °C 10 s, 62 °C 10 s, 72 °C 20 s for 40 cycles | 1.90 |
OPG | CTGCGCGCTCGTGTTTC ACAGCTGATGAGAGGTTTCTTCGT | 300 | 95 °C 30 s, 60 °C 60 s, 72 °C 15 s for 40 cycles | 1.99 |
RANK-L | CATCCCATCTGGTTCCCATAA GCCCAACCCCGATCATG | 300 | 95 °C 10 s, 60 °C 10 s, 72 °C 15 s for 40 cycles | 2.07 |
TNF α | CCCAGGGACCTCTCTCTAATC GCTTGAGGGTTTGCTACAACATG | 300 | 95 °C 30 s, 60 °C 60 s, 72 °C 15 s for 40 cycles | 1.97 |