Plasmodium falciparum (controls) Amplicon deep-sequencing using Pf-SMARRT panel,
Amplicon sequencing libraries were prepared using the PfSMARRTer protocol, a streamlined version of the previously published PfSMARRT assay for Plasmodium falciparum molecular surveillance. Genomic DNA extracted from P. falciparum isolates was used as input without normalization. The library preparation process involves a two-step PCR protocol: the first PCR amplifies a panel of targeted loci associated with antimalarial resistance (including full coverage of k13, mdr1, and dhfr), while the second PCR adds sample-specific barcodes and Illumina adapters. PCR products were pooled in 96-well plate format and purified using magnetic bead cleanup. The protocol is optimized for high-throughput, automation-compatible workflows and includes additional markers to detect emerging resistance mutations. Final libraries were quantified and sequenced using Illumina platforms.