Rajkumari Jashmi Devi, Merinashwari Konjengbam, Biseshwori Thongam
Chenopodium is one of the most complex genera in the family Amaranthaceae, exhibiting ploidy levels ranging from diploid to hexaploid. The lack of distinct morphological characters and phenotypic plasticity has complicated species identification in this genus. The advent of next-generation sequencing technology has enabled in-depth genomic studies of Chenopodium species. To date, the chloroplast genomes of six Chenopodium species-Chenopodium acuminatum, C. ficifolium, C. berlandieri, C. petiolare, C. quinoa, and C. album-have been reported using genomic DNA as templates. On the other hand, high-quality chloroplast DNA (cpDNA) is recommended for effective chloroplast genome sequencing. The present study reports an optimized high-salt extraction method for extracting cpDNA from C. album. The quality of isolated cpDNA was validated by determining the complete chloroplast genome of C. album using the Illumina platform with a 150 × 2-pair-end (PE) library and cpDNA as the template. The chloroplast genome of C. album is 152,028 base pairs and contains 129 genes. Of these, 84 are protein-coding genes, 8 are rRNA genes, and 37 are tRNA genes. This study lays the foundation for isolating high-quality cpDNA using a simple yet efficient method for extracting cpDNA from C. album, enabling efficient sequencing of the whole plastid genome for further understanding of the evolution, genetics, and taxonomy of this complex plant. © 2026 Wiley Periodicals LLC Basic Protocol: An optimized high-salt extraction protocol for chloroplast DNA isolation from Chenopodium album (Amaranthaceae), enabling whole plastid genome sequencing.