The study was conducted to evaluate and compare the effect of single and double dose prostaglandin on estrus response and conception of crossbred (Holstein Friesian x Zebu) and local (Zebu) cattle in Eastern zone of Tigray, Ethiopia. The study also evaluated and compared efficiency of AI technicians to detect cuprous luteum through rectal palpation and reproductive status by progesterone level determination in milk using Rapid Progesterone Heat Detection Test (RPHDT). A total of 240 crossbred postpartum (60 days) cows and heifers were assigned for single (group 1, n=120) and double (group 2, n=120) dose PGF2 protocols. Estrus response and conception rate was compared between protocols, breeds and parity. Reproductive stage of animals was confirmed prior to estrus synchronization and efficiency of AI technicians on detection of CL through rectal palpation assessed using RPHDT. Among cows/heifers synchronized using both PGF2 protocols, 87.2% (157/180) were detected in estrus on visual observation and rectal palpation. Overall conception rate among animals in heat was 62.7% (96/153).