Rnase free dnase 1 amplification grade kit

RNA samples were collected after infecting the primary brain cell cultures by the 76 K strain for 24 h, 48 h, 96 h, 7 days and 14 days. Uninfected brain cell culture samples were collected at the same time as the 24 h infected cells time point. Infected and uninfected cells were washed with 1 ml of PBS (two times) and lysed by a direct load of Trizol in the plate. RNA was extracted as per manufacturer instruction and genomic DNA was removed using the RNase-free DNase I Amplification Grade Kit (Sigma). All RNA samples were assessed for quality using an Agilent 2100 Bioanalyzer. RNA samples with an integrity score greater than or equal to 8 were included in the RNA library preparation. Triplicates (biological replicates) were produced for each condition. The TruSeq Stranded mRNA Sample Preparation kit (Illumina) was used to prepare the RNA libraries according to the manufacturer's protocol. Library validation was carried out by using DNA high-sensitivity chips passed on an Agilent 2100 Bioanalyzer. Library quantification was carried out by quantitative PCR (12 K QuantStudio).

RNA samples were prepared by infecting T175 flasks containing HFF cell monolayers with iKD TgAP2IX-5 parasite for 24 h followed by 6 h auxin treatment before sample collection and addition of Trizol (Invitrogen). Control samples were left to grow in normal media. Auxin was added for 6 h since this is the duration required for T. gondii tachyzoites to complete a full cycle of asexual division. RNA was extracted as per manufacturer instruction. This was then followed by genomic DNA removal and cleaning using the RNase-free DNase I Amplification Grade Kit (Sigma). All RNA samples were assessed for quality using an Agilent 2100 Bioanalyzer. RNA samples with an integrity score greater than or equal to 8 were included in the RNA library preparation. Triplicate (biological replicate) were produced for each conditions.