porcine T. foetus transcriptome (PIG30/1)
Porcine Tritrichomonas foetus (PIG30/1)were axenized from faecal samples of domestic pig and cultured through 6 passages in Modified Diamond's media, supplemented with 10% heat-inactivated sheep serum. Trophozoites at mid-exponential phase in culture and total RNA was isolated from 10*7 cells. RNA isolation was carried out using the RNeasy Micro kit (Qiagen), according to the manufacturer’s instructions. Homogenisation was carried out in a FastPrep® - 24 Instrument (MP Biomedicals, USA) for 30 seconds at 4 m/s. An in-column DNAase (Sigma-Aldrich) treatment step was included, which was at room temperature and an incubation time of 15 minutes. RNA was eluted in 30 μl of sterile water and assessed both, qualitatively and quantitatively, using a 2100 Bioanalyzer (Agilent Technologies, Inc). The sample with superior quality was preserved in a RNAstable® tube (Biometrica) by drying in a Savant SpeedVac concentrator connected to a vapour trap for 1 hour, in accordance with the manufacturer’s instructions. Paired-end RNA sequencing on an Illumina HiSeq2000 platform was performed at Macrogen (Seoul, Korea).