Allele frequencies and haplotype diversity of 17 Y-chromosomal short tandem repeat (STR) loci included in Y-filer™ PCR amplification kit were studied in 120 Garo and 139 Santal male individuals residing in two distinct regions of Bangladesh. A total of 99 different haplotypes from Garo and 129 different haplotypes from Santal individuals were observed with a corresponding discrimination capacity (DC) of 0.825 and 0.928, respectively. A comparison of the studied data with the published data from Y-STR haplotype reference database (YHRD) based on AMOVA revealed that the Garo population is closely related to Tripuri population from Tripura, India and Santal population moderately close to Munda population from Jharkhand, India. The mainstream Bengali population resides at a significant genetic distance from these two studied populations.