Figure 3

Relative var transcript levels in the pRBC of three distinct FCR3S1.2 cultures as seen by qPCR. The most frequently identified FCR3S1.2 var transcripts were amplified and the levels were compared to that of the endogenous control, Seryl-tRNA-synthetase. The var genes amplified had previously been identified by semi-quantitative PCR and sequencing by employing three sets of degenerate primers to the 5-prime end of the var genes[12]. Transcription levels were measured in different cultures of FCR3S1.2. A) 18 generations after micro-manipulation cloning (FCR3S1.2 18 G; 85% rosetting; [33], B) 100 generations after cloning (FCR3S1.2 >100 G, enrichment on Ficoll to maintain rosetting, 85% rosetting), and C) at 28 generations of additional growth of FCR3S1.2 100 G without Ficoll enrichment (FCR3S1.2 128 G, 78% rosetting). Results are visualized as log2 transformed values.