Comparison of three extraction methods in the preparation of good yield and quality of total RNA from Toxoplasma gondii

In this study, we evaluated three extraction methods for their abilities to extract a good yield and quality (purity and integrity) of total RNA from Toxoplasma gondii tachyzoites for cDNA library construction. The yield and purity of intact total RNA extracted was assessed using nanophotometer, while the integrity of the total RNA was evaluated using Agilent Bioanalyzer. The genomic DNA contamination of intact total RNA extracted and the success of cDNA synthesis were validated using Stratagene Easy-A® One Tube RT-PCR System. All three extraction methods were found to be suitable for extraction of the T. gondii total RNA. However, the precipitation-based RNA extraction method comparatively produced the highest yield of total RNA from approximately 1x10 6 parasites. The reagent was also found to stabilise and protect the total RNA in stored specimens from degradation. As our study targeted 3 mg of total RNA for cDNA library construction, therefore, the precipitation-based RNA extraction method appeared to be the most suitable method to extract a good yield and quality of T. gondii total RNA.