4. 7 Real-time PCR assays for quantification of parasites
Total RNA was isolated from the infected liver, and cDNA was synthesized as previously described (Choudhary et al. , 2019). The parasite burden was quantified by amplifying 18S rRNA (Bruña-Romero et al. , 2001) using primers 1195/1196. MSP1 transcripts were quantified using primers 1219/1220. Liver samples were normalized to mouse glyceraldehyde-3-phosphate dehydrogenase (GAPDH) using primers 1193/1194. To quantify the copy number, gene-specific standards were run alongside the unknown samples.