The authors designed six primers for the amplification of three non-coding regions of chloroplast DNA (cpDNA) via polymerase chain reaction (PCR). These primers were tested on DNA from various plant species, including algae, bryophytes, pteridophytes, gymnosperms, and angiosperms. The primers successfully amplified the target regions in most species, demonstrating their universality. The non-coding regions, which have a higher frequency of mutations, can be used to study population biology and evolution at both interspecific and intraspecific levels. The primers were designed to flank conserved regions, increasing their taxonomic range. The sequences of the primers and the PCR conditions are provided, along with the expected sizes of the PCR products. The study highlights the potential of these primers for genetic marker development and phylogenetic analysis in plants.The authors designed six primers for the amplification of three non-coding regions of chloroplast DNA (cpDNA) via polymerase chain reaction (PCR). These primers were tested on DNA from various plant species, including algae, bryophytes, pteridophytes, gymnosperms, and angiosperms. The primers successfully amplified the target regions in most species, demonstrating their universality. The non-coding regions, which have a higher frequency of mutations, can be used to study population biology and evolution at both interspecific and intraspecific levels. The primers were designed to flank conserved regions, increasing their taxonomic range. The sequences of the primers and the PCR conditions are provided, along with the expected sizes of the PCR products. The study highlights the potential of these primers for genetic marker development and phylogenetic analysis in plants.