Introduction: Paternity test is a DNA examination to determine whether a man is a biological father of a child through DNA pattern comparison between the child and the suspected father to verify DNA inheritance proof. Methods: This study was conducted through analysis of kinship analysis application on paternity test through STR CODIS examination in siblings using peripheral bloods samples of forty samples Madurese 3 generations. The examined locuses were CSF1PO, TH01, TPOX, vWA. Results: DNA samples isolates found that the mean level of DNA was 595.35 ± 4.15ng/µl, whereas the range of purity was 1.17 – 1.29. From this study, alleles from maduranese family population in Surabaya was variable, the highest percentage was CSF1PO locus allele 9 [33.75%], THO1 allele 8 [26.25%], TPOX allele 9 [32.5%] and vWA allele 17 [21.25%]. This study also revealed shared alleles on one sibling allele, the highest was 2 shared alleles on CSF1PO [70%], THO1 [100%], TPOX [70%] and vWA locus [90%]. Conclusion: THOI locus had 100% 2 shared alleles so that it was recommend using STR locus to identify kinship on Madurese in Surabaya.