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Fig. 1 | Malaria Journal

Fig. 1

From: Improved Plasmodium falciparum dilution cloning through efficient quantification of parasite numbers and c-SNARF detection

Fig. 1

Schematic representation of steps using C-SNARF detection for P. falciparum cloning limiting dilution. In step 1, 5 µl of the medium-depleted cultured red blood cells are removed and diluted in 995 µl RPMI, and the parasitaemia is determined by flow cytometry using DNA stain EtBr or equivalent (step 2 and 3). After calculation of the total number of number of parasites per µl, a microliter volume corresponding to 20 parasites (dilution may be done stepwise) is mixed to result in 9.5 ml of a complete medium to which 0.5 ml of fresh blood is added (step 4). In sequence, 100 µl of this suspension is dispensed in 96 wells and the plate is then kept under the appropriate atmosphere and medium is changed on day 7, increased on day 11 to 200 µl and on day 14, the medium is changed to HEPES-free medium plus C-SNARF (step 5). At day 16 and 18 positive wells are detected by C-SNARF fluorescence (step 6)

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