These bands appear in new positions, because denatured DNA has a higher density than native DNA.

In semiconservative replication, the single strands of denatured DNA should have half the molecular weight of the native hybrid molecule. The experimental results favored again the semiconservative model, but the experimental accuracy was not sufficiently high to make the results entirely clear-cut. However, put together, the evidence in favor of semiconservative replication is sufficient to allow us to accept this model. In this connection (see also later) we should mention that the denatured DNA can reform to the native from in spite of the fact that in denatured DNA the two strands have come apart. We note in passing that there is relatively little DNA between three band positions indicating that the extracted DNA has almost always the densities of pure N15-N15, pure N14-N15, or pure N14-N14 DNA. this observation implies that the replication process replicates a complete length of DNA before going on to the next molecule. However, this dramatic conclusion applies only to the relatively short length of DNA which were prepared for analysis by the methods available at the time of the Messelson-Stahl experiment. Had the molecules survived intact, then such unsynchronized cell populations would have produced some DNA of intermediate buoyant densities, and this would have obscured the resultant picture. PROPERTIES OF DNA in the isolation of DNA a major difficulty is to keep such a long rigid molecule physically intact. it has been shown that the shear forces set up when a solution of DNA is vigorously stirred are sufficient to break the long rigid DNA molecule. The molecules are first broken near the middle, then into quarters, and so on until a short-enough size is reached that the molecule will be relatively insensitive to breakage by shearing. It is difficult to extrapolate the molecular weight of DNA found in solution to the actual molecular weight that exists in vivo because of the great ease of degradation by shearing during extraction or by the inadvertent action of hydrolytic enzymes. perhaps the best estimates are derived from radioautographic work in wich the DNA is labeled with tritium. for example, in E coli, cairns was able to detect DNA molecules up to 1.1 mm in length. this corresponds to a molecular weight of 2.8x109, similarly the DNA from T2 bacteriophage has an average length of 49 micras, wich would correspond to a molecular weight of 108. a rather smaller bacteriophage, called lambda, has a DNA 23 micras in length. but even in the most carefully prepared samples of extracted DNA there is always 0.1 to 0.2 percent of protein attached to the material. therefore the possibiliy existed that a DNA molecule is actually made up shorter lengths of DNA held together by protein peptide-like materials. however, the successful synthesis in vitro by known reactions of the circular doublestranded replicative form of 174 DNA, capable of infecting host cell, makes it likely that such DNA molecules are composed only of nucleotides.