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.