The red algal family Gracilariaceae is commercially valuable due to its use in biotechnology and microbiology research as a phycocolloid agar. Marine macroalgae, especially the Rhodophyta, can be notoriously difficult to identify owing to their relatively simple morphology and anatomy, convergence, rampant phenotypic plasticity, and alternation of heteromorphic generations. The monophyly of the red algal family Gracilariaceae has been confirmed by 18S rDNA sequence analyses. We tested the effectiveness of DNA barcoding in the identification and discovery of Gracilariaceae species. Our analysis of 18S rDNA sequence yielded a three species namelyGracilaria, Grateloupia, and Hypnea. One locus was tested for their use as DNA barcodes in three red algae. A fragment of 1552, 1547 and 1556 bp of 18S rDNA region was analyzed in three specimens namely Gracilaria, Grateloupia and Hypnearespectively.Based on the present study it can be concluded that the above data provide strong evidence that Gracilaria, Grateloupia, and Hypnea are distinct evolutionary entities and should be recognized as separate genera. Our study demonstrates that DNA barcoding can provide an efficient method for species-level identifications and contribute powerfully to taxonomic and biodiversity research.