Steril rnase free water

Total RNA was extracted from frozen cell pellets with TRIzol™ Reagent (Life Technologies, Carlsbad, CA, USA) and the following chloroform phase separation and ethanol precipitation at −20 °C. Washed RNA was dissolved in Steril RNAse-free water (Thermo Fisher Scientific, Waltham, MA, USA). The final RNA concentration was measured using a spectrophotometer. To prevent RNA aggregation, total RNA samples were heated at 65 °C for 1–2 min before cDNA synthesis; samples were immediately used in cDNA synthesis. DNAse treatment was performed with RQ1 RNAse-free DNAse (Promega, Madison, WI, M6101). First-strand cDNA was synthesized from 1 µg total RNA using Mint Kit (Evrogen, Moscow, Russia) according to the manufacturer’s instructions.

Total RNA was extracted from defrosted and homogenized SH-SY5Y cells with TRIzol™ Reagent (Life Technologies, Carlsbad, CA, USA) and the following chloroform phase separation and ethanol precipitation at −20 °C. Washed RNA was dissolved in Steril RNAse-free water (Thermo Fisher Scientific, USA). Final RNA concentration was measured using a spectrophotometer. To prevent RNA aggregation, total RNA samples were heated at 65 °C for 1–2 min before cDNA synthesis. First-strand cDNA synthesis was performed using Mint Kit (Evrogen, Moscow, Russia) according to the manufacturer’s protocol (1.5 μg RNA for each reaction).