TaqMan RT-PCR for miRNA expression

All reagents including specific and control primers for TaqMan real-time RT-PCR were purchased from Applied Biosystems and the reactions were set according to the manufacturer’s protocol. Briefly, total RNA was purified by TRIzol (Life Technologies/Invitrogen). 10 ng of total RNA and RT primers for hsa-miR-132, hsa-miR-212 and the housekeeping gene RNU44 were used for reverse transcription (RT) with TaqMan MicroRNA Reverse Transcription Kit. Real-time PCR quantification was performed using TaqMan PCR primers and TaqMan Universal PCR Master Mix, No AmpErase UNG on the ABT PRISM 7900HT Fast PCR system (Applied Biosystems). Relative mRNA expression was determined as previously described [26] (link).