Random decaprimer

Tissue samples were collected in 1 h after the start of stimulation, placed in 1.5 mL tubes, and frozen in liquid nitrogen. RNA isolation was performed using an ExtractRNA reagent (Evrogen, Moscow, Russia) in accordance with the manufacturer’s recommendations. To remove traces of genomic DNA, RNA samples were treated with DNase I (Thermo Scientific, Vilnius, Lithuania). Reverse transcription was performed using the MMLV RT reagent kit (Evrogen, Moscow, Russia) and murine RNase Inhibitor (New England Biolabs, Ipswich, MA, USA) as recommended by the manufacturer. An equimolar mixture of random decaprimer (Evrogen, Moscow, Russia) and oligo(dT)15 primer (Evrogen, Moscow, Russia) was used; the concentration of each primer in the reaction was 1 μM. After reverse transcription, the reaction mixture was diluted 8-fold with deionized water.

In all experiments, tissue samples were collected 1 h after the start of stimulation, placed in 1.5 mL tubes, and frozen in liquid nitrogen. RNA isolation was performed using an ExtractRNA reagent (Evrogen, Moscow, Russia) in accordance with the manufacturer’s recommendations. To remove traces of genomic DNA, the RNA samples were treated with DNase I (Thermo Scientific, Vilnius, Lithuania). Reverse transcription was performed using the MMLV RT reagent kit (Evrogen, Moscow, Russia) and murine RNase Inhibitor (New England Biolabs, Ipswich, MA, USA) as recommended by the manufacturers. An equimolar mixture of random decaprimer (Evrogen, Moscow, Russia) and oligo(dT)15 primer (Evrogen, Moscow, Russia) was used, and the concentration of each primer in the reaction was 1 μM. After reverse transcription, the reaction mixture was diluted eightfold with deionized water.