Complementary combination of multiplex high‐throughput <scp>DNA</scp> sequencing for molecular phylogeny

The rapid development of DNA sequencing technology in recent years has provided new tools for phylogenetic data acquisition. By using high-throughput technology, molecular information can be obtained more quickly and economically. Here, we describe a complementary combination two multiplex methods. One is multiplexed marker (MPM-seq), the other inter-simple sequence repeat (ISSR) genotyping by (MIG-seq), whose protocol improved over that original one. Both MPM-seq MIG-seq begin with polymerase chain reaction (PCR), each amplifying multiple markers genome-wide ISSR regions, respectively. After another PCR second primer set common both methods, next-generation used to simultaneously detect sequences regions from samples method. In this case study, performed analysis Japanese fir (Abies) closely related Abies species. revealed three chloroplast one nuclear internal transcribed spacer created partially informative tree 13 Whereas detected 6700 single-nucleotide polymorphisms exhibited clear clustering species 97%–100% bootstrap support all branches tree. Hence, combination, quick, simple, economical wide range genomic studies, including phylogeny, as well investigating genetic differentiation or identification among species, hybrids, populations, even clones cultivars, barcoding technique.