DNA and RNA Extraction Protocol

Exponential cell cultures (1-3 × 10⁵ cells/ml) were harvested by centrifugation at 2,800 rpm for 3 min. Total DNA was isolated using the protocol described in Martín-Platero et al., (2007) (link) and samples were treated with 10 mg/ml RNase A (Thermo- Scientific) for 2h at 37°C. Total RNA samples were isolated by using the TRIzol Reagent method (Invitrogen). RNA samples were treated with DNase I (Roche) for 30 min at 37°C. Both, DNA and RNA integrities were tested by agarose gel electrophoresis. DNA or RNA sample concentrations were calculated by the NanoDrop 1000 (Thermo Scientific). MultiScribe Reverse Transcriptase 50 units/μl (Life Technologies) and oligo(dT)-adaptor primer (Roche) were used to synthesize cDNAs from 3.5 μg of total RNA.