Megaplex rt primers human pool a and b

Reverse transcription of isolated miRNA from ten laryngeal squamous cell carcinoma, patient blood samples and ten controls was performed according to the manufacturer’s recommendations using Megaplex™RT Primers Human Pool A and B, TaqMan® Human MicroRNA Array A and B purchased from Applied Biosystems using a 7900 HT System (Applied Biosystems). The expression levels of 377 human miRNA genes for each group were analysed. The following miRNAs revealed an altered expression profile: miR-122, miR-21, miR-155, miR-222, miR-181a, LET-7a, miR-31, miR-141, miR-149, miR-182, miR-145, miR-223, miR-133a, miR-485, miR-122, miR-33. These were chosen for further investigations.

miRNA expression was first screened using TaqMan^® Human MicroRNA Array A and B (Applied Biosystems, Foster City, CA, USA). Megaplex™RT Primers Human Pool A and B were purchased from Applied Biosystems and used to prepare the reverse transcription reaction according to manufacturer’s instructions. Amplifications were performed on a 7900HT Fast Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) under the following conditions: 2 min at 50 °C, 10 min at 94.5 °C, and 40 cycles each for 30 s at 97 °C and 1 min at 57 °C. The expression levels of 754 human miRNA genes were assessed in the first step in the screening cohort of randomly chosen FFPE endometriosis samples (OE) (n = 20) and controls (CG) (n = 20) and in ovarian cancer samples (n = 20 for high-grade ovarian cancer + 20 for endometrioid ovarian cancer). miRNAs with altered expression profiles (based on 2-fold change, PCR amplification curves, and confirmed in the literature) were chosen for further investigations in the examined groups of patients: DIE, SE, OE, and CG.