Table 2.
Comparison of expected and observed band sizes from Southern blot analysis using the dhodh probe
| Resistance level | Clone | Average # of amplicons from qPCRa | Predicted increase in size b (Mb) | Expected size (Mb) | Observed size (Mb) |
|---|---|---|---|---|---|
| Wild type | WT1 | 1 | — |
1.4 c |
~1.8 |
| WT2 | 1 | — | 1.3 c | <1.8 | |
| Low | L1 | 4 | 0.292 | ~2.1 | 1.8–2.4 |
| Moderate | M1 | 8 | 0.584 | ~2.4 | 1.8–2.4 |
| High | H1 | 10 | 0.730 | ~2.5 | 1.8–2.4 d |
| H4 | 12 | 0.876 | ~2.6 | 1.8–2.4 |
a
These values were derived from qPCR on purified genomic DNA and previously presented in (Guler et al., 2013). They likely represent the total number of amplicons present per parasite (chromosomal + ecDNA).
b
Number of amplicons (determined by qPCR) x amplicon size (Guler et al., 2013).
c
The size of WT1 (Dd2) and WT2 (Hb3) chromosome 6 was retrieved from PlasmoDB (Aurrecoechea et al., 2009).
d
H1 chromosome 6 appears smaller than that from M1, despite an increased number of amplicons. Next Generation Sequencing revealed a ~10 kb deletion on chromosome 6 in a related clone (H2, Guler et al., 2013), but this does not account for the size difference.