4.3.2. Gene expression of osteogenic differentiation markers by RT‐PCR

Total RNA was extracted from hMSCs seeded and cultured on plate control, CaSO₄, β‐TCP, β‐TCP‐HA, L‐RSF and M‐RSF on Days 7 and 28 by TRIzol method (Invitrogen Life Technologies, Carlsbad, CA) as mentioned above. Concentration of RNA was measured using Nano‐drop. cDNA was prepared from 200 ng of total RNA using Verso cDNA synthesis kit according to manufacturer's instructions. The PCR conditions used and annealing temperatures for each gene are as described in Table S3. PCR products were resolved on 1.2% agarose gel and visualized using SYBR gold stain (Invitrogen) on Bio‐Rad, Molecular Imager, ChemiDox™ XRS+ imaging system. Band intensity was then analyzed using ImageJ software. Values were normalized using GAPDH (housekeeping gene control).