The present study was carried out in the period May-August 2022 in An-Najaf Province, Iraq. The study aimed to classify hard ticks in cattle through molecular techniques. About 468 hard ticks were collected from naturally infested cattle which considered an important economic source of income. Phenotypic examination, then Polymerase Chain Reaction (PCR) were applied using a primer for cox1 gene for the diagnosis of Rhipicephalus. Sequencing of 18S rRNA gene was done to identified the Rhipicephalus species. Result of Rhipicephalus genus was proven in PCR, while sequence analysis and phylogenetic tree appeared two species Rhipicephalus turanicus and Rhipicephalus glabroscutatum that was recorded for the first time in Najaf.