Meselson-Stahl showed the semi-conservative mechanism of DNA replication. They cultured E. coli cells initially in a medium containing ammonium salts prepared with heavy nitrogen 15N15 to label, using all cellular DNA. Then cells were transferred to a medium containing the normal light isotope 14N14. DNA was analyzed by equilibrium density-gradient centrifugation in the periodically collected sample to separate heavy-heavy (H-H), light-light (L-L), and heavy-light (H-L) duplexes into distinct bands. The sample was placed on a preformed gradient of cesium chloride. Centrifugation separates the macromolecules in different bands of density gradient which can be collected as a pure fraction. After the first generation, the density gradient contained only one band having H-L nitrogen. After the second generation, the density gradient contained 2 bands- one will have hybrid DNA (H-L) and the other will have light DNA (L-L).
Thus, the correct answer is option D.