Megaplex rt primers specific for each card

The miRNA expression profiling was performed by the TaqMan^® Array Human MicroRNA A+B Cards Set v3.0 (Life Technologies, USA) containing a total of 384 miRNA probes and controls per card. Overall, we analyzed 15 FF samples and 15 FFPE samples (five HPV-positive tumor samples, five HPV-negative tumor samples, and five non-malignant tissues of each material). For the analysis of FF samples, 1000 ng of total RNA was used for reverse transcription. In the case of FFPE samples, the input was 350 ng due to the smaller concentration of extracted RNA from FFPE samples. Total RNA of each sample was reverse transcribed using the TaqMan^® MicroRNA Reverse Transcription Kit and Megaplex^™ RT Primers specific for each card (both Life Technologies, USA) according to the manufacturer's protocol. The workflow without preamplification was chosen for both types of material, FF as well as FFPE samples. The TLDA cards were analyzed on the Applied Biosystems 7900HT Real-Time PCR System.

The global quantitation of miRNA gene expression was performed using the TaqMan® Array Human MicroRNA A + B Cards Set v3.0 (Life Technologies, USA) containing a total of 384 TaqMan® MicroRNA Assays and controls per card. Each array contains three assays of endogenous controls (RNU48, RNU44, and RNU6B) and one assay as a negative control.
Overall, we analyzed 35 clinical samples and 12 cell lines. First, 1000 ng of total RNA of each sample were reverse transcribed using the TaqMan® MicroRNA Reverse Transcription Kit and Megaplex™ RT Primers specific for each card (both Life Technologies, USA) according to the manufacturer's instructions. The TLDA cards were analyzed on the Applied Biosystems 7900HT Real-Time PCR System.