Fam dye labeled mgb probes

Total RNA or microRNA from cultured cells were extracted with mirVana miRNA Isolation Kit (Ambion) according to the manufacturer’s instructions. Reverse transcription of mRNA was performed using the High Capacity RNA-to-cDNA kit (Applied Biosystems). TaqMan MicroRNA Reverse Transcription Kit was used to produce cDNA for TaqMan MicroRNA Assay (Applied Biosystems). Real-time PCR was performed on the 7900HT Fast Real-Time PCR System (Applied Biosystems) using the TaqMan® Universal Mastermix (Applied Biosystems). Human and mouse CDC6 expression was quantified in real-time with CDC6-specific FAM dye-labeled MGB-probes and normalized to GAPDH (Applied Biosystems). Human miR-3178 was quantified in real-time with FAM dye-labeled MGB-probes and normalized to RNU66 or snU6 (Applied Biosystems).