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, Reverse transcription was performed with 500 ng of total RNA per sample using TagMan reverse transcription reagents (Applied Biosystems, Foster City, CA) with random hexamer primers. Gene detection was carried out by using specific oligonucleotide primers and TaqMan probes (Microsynth, Switzerland) and Assays on Demand (Applied Biosystems, Foster City, CA) (set at 95°C for 10 min, followed by 40 cycles of amplification at 95°C for 15 sec and 60°C for 1 min. The ribosomal protein L13a (RPL13A) was used as endogenous control, Total RNA was extracted using TRI-reagent® (MRC)

, Probe:5'-CAT CAA ACA GCC TCT TCA GCA CAG TGA CAC-3' Human collagen type I (hCol I) Forward

, Primer and probe sequences used for real-time polymerase chain reaction. 3.4.3.7. Mineralisation assessment by alizarin Red S staining

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