<p>Short tandem repeat (STR) markers are well established as the gold standard and boon for forensic DNA applications. India is well known for its population diversity. Since several population genetic studies have been undertaken, more diverse populations need to be explored. In this study, we first evaluated randomly selected 297 unrelated individuals belonging to the sikh population of Sri Ganganagar Rajasthan through 24 autosomal STR loci included in the Ingenomics™ AutoProfiler STR Kit. A wide range of allelic diversity was observed, with a total effective allele count of 6–41 among all the studied loci. Allele 8 of the TPOX locus was the most frequent allele in the studied population, with a value of 0.444. Locus SE33 was found to be the most useful locus in the studied set of markers for the studied population, with the highest number of effective alleles (Ne = 41), highest value of power of discrimination (PD = 0.991), polymorphic information content (PIC = 17.82), typical paternity indexTPI = 6.75), power of exclusion (PE = 0.849), highest number of heterozygotes (92.59%), lowest value of matching probability (pm = 0.009) and lowest number of homozygotes (741%). All the tested loci were useful together with the highest value of the combined power of discrimination (CPD = 1), the combined power of exclusion (CPE = 1), the combined paternity index (CPI = 1.37 × 10¹⁰) and the lowest value of the combined matching probability (CPM = 6.53 × 10^− 31). In the population comparison, the studied population showed genetic affinity with the populations of Rajasthan and Punjabi in Pakistan. This study used a population dataset for forensic DNA application and genetic studies.</p>