This dataset comprises Y-STR profiles and Y-SNP haplogroups of 327 unrelated Thai male individuals from the borderland provinces of Tak (n = 274) and Ranong (n = 53), Thailand. Genomic DNA was extracted from blood samples stored on Whatman™ FTA™ cards and analyzed using advanced genotyping techniques. Y-SNP haplogroup assignments were performed using yHaplo v1.1.2 and referenced against the ISOGG Y-DNA Haplogroup Tree v15.73. Y-STR profiling was conducted with the ForenSeq™ DNA Signature Prep Kit on the MiSeq FGx platform, ensuring high-resolution allele characterization validated in the Thai population.