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DNA replication copies a double-stranded DNA molecule into two daughter double-stranded DNA molecules. In semiconservative replication, each daughter retains one parental strand and receives one newly synthesized complementary strand. DNA polymerases build the new strands from incoming nucleotides; they do not make RNA or proteins in this copying step. See Alberts et al., Molecular Biology of the Cell, 4th edition (2002): DNA replication mechanisms.

For a five-base pair example, separate $5^{\prime }\text{-}\mathrm{ATGCC}\text{-}{3}^{\prime }$ from its partner $3^{\prime }\text{-}\mathrm{TACGG}\text{-}{5}^{\prime }$. Base pairing specifies a new TACGG partner for the old ATGCC strand and a new ATGCC partner for the old TACGG strand. With error-free copying, both daughter DNA molecules have the original nucleotide sequence, but different physical strand histories. One initial duplex contains ten nucleotides; one complete round produces two duplexes containing twenty nucleotides, ten retained and ten newly incorporated.

This is useful for inheritance: both daughter biological cells can receive a copy of the DNA record. The elegant feature is that base pairing lets each strand specify its missing partner. Direction matters too: a replicating DNA polymerase reads its template $3^{\prime }\to 5^{\prime }$ and extends the new strand at its $3^{\prime }$ end, so synthesis proceeds $5^{\prime }\to 3^{\prime }$. These are opposite directions on antiparallel nucleic acid strands, not two alternative directions of synthesis. The enzyme needs a pre-existing paired strand end to extend; the diagram omits the machinery that starts and coordinates copying. See Alberts et al.: template-directed synthesis and the requirement for a primer.

The strand accounting predicts observable differences between copying mechanisms. In 1958, Meselson and Stahl grew Escherichia coli with the heavier nitrogen isotope ${}^{15}\mathrm{N}$, then supplied ${}^{14}\mathrm{N}$ and measured extracted DNA by density-gradient centrifugation. After one generation their samples contained intermediate-density DNA; after two, intermediate-density and light DNA occurred in equal amounts. This supported semiconservative replication in the tested Escherichia coli culture. It was an isotope assay of sampled DNA, not simultaneous testing of every kind of DNA or direct colored-strand photography. See The replication of DNA in Escherichia coli by Matthew Meselson and Franklin W. Stahl.

In isopycnic centrifugation, a species concentrates near the position where the liquid's density matches its buoyant density. In a density gradient increasing outward, material displaced inward encounters less-dense liquid and tends to move outward; material displaced outward encounters denser liquid and tends to move inward. Diffusion broadens the resulting band. If the density gradient never reaches the species' buoyant density, a matching equilibrium position need not exist within the vessel.^[ref]

The 1958 paper The replication of DNA in Escherichia coli by Matthew Meselson and Franklin W. Stahl (pp. 672–676, Figures 1–4) makes the distinction concrete. Its separation apparatus was a Spinco Model E ultracentrifuge with an Escherichia coli lysate in a cesium chloride density gradient. Ultraviolet wave absorption photographs of the rotating sample were scanned with a recording microdensitometer: peak position reported buoyant density, while signal above baseline reported DNA concentration.^[ref]

Mixed lysates labeled fully with the nitrogen isotope ${}^{15}\mathrm{N}$ and unlabeled lysates supplied reference bands; a mixture including intermediate material checked its position relative to those references. Figure 1 followed band formation over time, helping distinguish a migrating distribution from an approximately stationary one. These controls address resolution, calibration and approach to equilibrium. They do not make every ultraviolet-absorbing component identifiable as DNA, or turn a band into a direct image of individual strands. The DNA replication article develops the copying inference; here the key result is a separation by buoyant density, not a claim that heavier-labeled DNA is physically larger.^[ref]