The interaction of the glutamic acid (Glu) with double-stranded (ds) calf thymus DNA (CT-DNA) has been studied by absorption and emission spectroscopy then further results are compared with molecular docking study. The calculated value of binding constant estimated by different techniques are very close to each other, which is around 3.54×103 ML-1. Besides, the value of binding sites from fluorescence is 0.939, which suggests appropriate binding of Glu with CT-DNA. The result shows Glu binds to nitrogenous bases (adenine and guanine) of CT-DNA through the groove binding mode of interaction. The experimental results were compared successfully with docking results. The binding energy (ΔG) of the complex has been -4.76 kCal M-1 and -5.1 kCal M-1 from fluorescence and molecular docking studies, respectively. In our study a correlative intractability pattern for Glu with DNA has been identified.