Table 1 |
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Description of the LNA probe and primer sequences used to quantify gene expression |
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|
Gene |
Gene ID |
Sequences |
Product length |
Efficiency |
|
|
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|
iNOS |
4843 |
Probe: 5'-FAM-ACTTCTTTCCCGTCTCC-BHQ1-3' |
305 |
1.99 ± 0.8 |
|
Sense: 5'-TCCAGATAAGTGACATAAGTG-3' |
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|
Antisense: 5'-CAGCTTGACCAGAGATTC-3' |
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|
COX-2 |
5743 |
Probe: 5'-AAACTGCTCAACACCG-BHQ1-3' |
216 |
1.99 ± 2.8 |
|
Sense: 5'- GGACAGGATTCTATGGAG-3' |
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|
Antisense: 5'- GGATGTCAACACATAACTC-3' |
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|
Aggrecan |
176 |
Probe: 5-'FAM-CCAACTCTTCAAGGTGA-BHQ1-3' |
109 |
1.98 ± 0.4 |
|
Sense: 5'-GACTGAAGTTCTTGGAGAA-3' |
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|
Antisense: 5'-CACGAAAACCCAGAGTAA-3' |
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|
Collagen type II |
1280 |
Probe: 5'-FAM-TCTGTCTCCTTGCTTGCCA-BHQ1-3' |
200 |
1.99 ± 0.9 |
|
Sense: 5'-GGAGTCAAGGGTGATCGT-3' |
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|
Antisense: 5'-CTTGTGCACCAGCTTCTC-3' |
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|
GAPDH |
2597 |
Probe: 5'-HEX-CAGTCAGCCGCATCTTCT-BHQ1-3' |
160 |
1.98 ± 4.3 |
|
Sense: 5'-TCTCTGCTCCTCCTGTTC-3' |
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|
Antisense: 5'-CGCCCAATACGACCAAAT-3' |
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|
|
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|
Primers used in PCR experiments with Locked Nucleic Acid (LNA) probes produced amplicons between 109 and 305 base pairs and efficiency values between 1.98 and 1.99. Probes contain fluorescein (FAM) or 6-carboxyhexafluorescein (HEX) as the 5'-reporter dye and Black Hole Quencher 1 (BHQ1) as the 3'-quencher. Nucleotides highlighted in bold denote the LNA base. |
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|
Ramachandran et al. Arthritis Research & Therapy 2011 13:R145 doi:10.1186/ar3459 |
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