A method of identifying parentage in Macrobrachium rosenbergii prawn comprising the steps of amplifying targeted EST-SSR nucleic acid sequences from a biological sample of Macrobrachium rosenbergii through a polymerase chain reaction using primer pairs with nucleic acid sequences as set forth in SEQ ID NO. 1 and SEQ ID NO. 2, SEQ ID NO. 3 and SEQ ID NO. 4, SEQ ID NO. 5 and SEQ ID NO. 6, SEQ ID NO. 7 and SEQ ID NO. 8, SEQ ID NO. 9 and SEQ ID NO. 10, SEQ ID NO. 11 and SEQ ID NO. 12 to produce respective amplicons; determining fragment size of the produced amplicons; regarding the fragment size of the amplicons corresponding to predetermined alleles; and identifying possible parentage of the prawn based on the regarded allele.