Benih ikan baramundi Lates calcarifer diperoleh dari pemijahan alami dengan jumlah induk terbatas sehingga variabilitas pertumbuhan dan kelangsungan hidup antar-batch menjadi tinggi. Penelitian dilakukan untuk menganalisis pertumbuhan dan mengevaluasi variasi genetik ikan kakap putih populasi Australia, Situbondo dan Lampung hasil domestikasi dan dibudidaya di hatcheri skala rumah tangga (HSRT). Sebanyak 10 ekor ikan barramundi dari setiap populasi digunakan untuk analisis variabilitas genetik dengan dua lokus mikrosatelit, yaitu Lca21 dan Lca32. Selanjutnya, data mikrosatelit diolah menggunakan software genetic analysis in excel (GenAlEx 6.51b2). Hasil yang diperoleh menunjukkan bahwa pertumbuhan panjang dan bobot tubuh serta laju pertumbuhan spesifik ikan baramundi dari Australia lebih tinggi dibandingkan dari Situbondo dan Lampung (P<0,05); sedangkan ikan baramundi dari Situbondo dengan Lampung adalah sama (P>0,05). Jumlah alel setiap lokus ikan baramundi berkisar 2-8 alel dan heterozigositas tertinggi dimiliki oleh ikan barramundi asal Situbondo (0,85), diikuti Lampung (0,65) dan paling rendah dari Australia (0,54). Dari hasil riset tersebut dapat disimpulkan bahwa ketiga populasi ikan baramundi hasil domestikasi dan dipeliharan dalam sistem HSRT memenuhi kelayakan untuk digunakan untuk kegiatan hibridisasi atau membentuk populasi awal (sintetis). Barramundi seeds, Lates calcarifer are usually sourced from natural spawning using a limited number of broodstock. Therefore, the growth and survival rate of these seeds vary greatly between batches. The research was performed to determine the growth pattern and genetic variations of barramundi seed populations produced from domesticated broodstock sourced from Australia, Situbondo, and Lampung and reared in small-scale hatcheries. Ten individuals of barramundi from each population were used for microsatellite analysis using two microsatellite loci, namely: Lca 21 and Lca 32. The resulted microsatellite data was processed using the genetic analysis available in Excel software (GenAlEx 6.51b2). The results showed that the growth in length and body weight as well as the specific growth rate of barramundi seeds produced from Australia broodstock were higher than that of Situbondo and Lampung (P<0.05) while the later two were similar (P>0.05). The number of microsatellite alleles ranged from 2-8 and the highest heterozygosity was obtained by barramundi seeds produced by Situbondo (0.85), followed by Lampung (0.65), dan Australia (0.54) broodstock. From the results of the research, it can be concluded that the three populations of barramundi fish, which were domesticated and reared in the HSRT system, meet the criteria for use in hybridization program or for forming a synthetic population.