Purelink rna mini kit with on column dnase set

Total RNA was isolated from up to 10 mg of tissue using PureLink™ RNA Mini Kit with on-column DNase Set (Invitrogen); tissue was homogenized in RNA lysis buffer using TissueLyser (Qiagen, Hilden, Germany; 2 × 2 min at 30 Hz; 5 mm stainless steel beads). Concentration and purity of isolated RNA was measured with NanoDrop™ spectrophotometer (Thermo Fisher Scientific); RNA integrity was assessed using 2100 Bioanalyzer instrument (Agilent Technologies, Santa Clara, CA). Purified RNA was stored in nuclease-free water at − 80 °C until library preparation.

Twenty samples were selected for transcriptomic analysis and assigned to the following 4 groups: adult, fresh tissue (AF, n = 5), adult, vitrified/warmed tissue (AV, n = 5), juvenile, fresh tissue (JF, n = 5) and juvenile, vitrified/warmed tissue (JV, n = 5; Fig. 1). Total RNA was isolated from up to 10 mg of tissue using PureLink^™ RNA Mini Kit with on-column DNase Set (Invitrogen); tissue was homogenized in RNA lysis buffer using TissueLyser (Qiagen, Hilden, Germany; 2 × 2 min at 30 Hz; 5 mm stainless steel beads). Concentration and purity of isolated RNA was measured with NanoDrop^™ spectrophotometer (Thermo Fisher Scientific, Waltham, MA); RNA integrity was assessed using 2100 Bioanalyzer instrument (Agilent Technologies, Santa Clara, CA). Purified RNA was stored in nuclease-free water at -80 °C until library preparation.