A novel series of 2-hydroxy pyrimidines (4a-l) were synthesized via (E)-thienyl chalcones (3a-l). (E)- thienyl chalcones were prepared by the reaction of thiophene-2-aldehyde (1) with various substituted acetophenones (2) in presence of NaOH in alcohol medium. Treatment of these chalcones (3a-l) with urea gave the title compounds 2-hydroxy Pyrimidines (4a-l). The structures of the newly synthesized compounds were assigned on the basis of IR, 1H-NMR, Mass spectral studies and elemental analysis. All the final synthesized compounds were evaluated for their In-Vitro antibacterial, antifungal and antitubercular activities. Some of the compounds have exhibited promising antibacterial, antifungal and antitubercular activities.