Expression of 8 genes was measured to confirm the RNA-seq results on the Rotor-Gene qPCR system (QIAGEN). Gene levels transcripts were evaluated with serial dilutions of plasmids (GenScript). Results were normalized with a housekeeping gene expression and accounting for the parasite’s developmental stage (Supplementary Method ). var genes expression was assessed by 3 independent methods. Published DBLα primers [12 (link)] and newly developed primers targeting the ATS domain (ATS-2 primers based on varDB dataset [31 (link)], Supplemental Data 2 ) were used on the Rotor-Gene qPCR system (QIAGEN) following the same methodology. The qPCR-TaqMan developed by Hofmann et al [32 (link)] targeting the ATS domain (ATS-1) was performed on a Viia7 qPCR system (Applied Biosystems) (Supplementary Method ).
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Confirming RNA-seq with qPCR Profiling
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Corresponding Organization : Institut de Recherche pour le Développement
Other organizations : Inserm, Centre National de la Recherche Scientifique, Hôpital Bichat-Claude-Bernard, Assistance Publique – Hôpitaux de Paris, University of Calgary, Université Paris Sciences et Lettres, Institut Curie, Institut d’Epidémiologie Neurologique et de Neurologie Tropicale, Université de Toulouse, Pharmacochimie et Pharmacologie pour le Développement, Université Toulouse III - Paul Sabatier
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Variable analysis
independent variables
- Expression of 8 genes
- Var genes expression
- Gene levels transcripts
- Var genes expression
- Normalized with a housekeeping gene expression
- Accounting for the parasite's developmental stage
- Serial dilutions of plasmids (GenScript)
- Published DBLα primers [12]
- Newly developed primers targeting the ATS domain (ATS-2 primers based on varDB dataset [31])
- QPCR-TaqMan developed by Hofmann et al [32] targeting the ATS domain (ATS-1)
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