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5-methyl-2-(pyridin-3-yl)-1,3-thiazole-4-carbohydrazide (<b>1</b>) on treatment with 4-fluorobenzaldehyde in presence of catalytic amount of acetic acid, accessed the target compound (<b>2</b>) with the yield of 79%. The target compound was confirmed by ¹H-NMR, ¹³C-NMR, FT-IR and LCMS. In vitro antibacterial activity against <i>Staphylococcus aureus</i> (<i>S. Aureus</i>), <i>Bacillus subtilis</i> (<i>B. subtilis</i>), <i>Escherichia coli</i> (<i>E. coli</i>), and <i>Pseudomonas aeruginosa</i> (<i>P. aeruginosa</i>) were carried out and compound <b>2</b> showed promising activity against <i>B. subtilis</i>. In addition, compound <b>2</b> was analyzed for DNA binding study. It revealed that compound <b>2</b> has a promising affinity towards DNA double helix.