The elucidation of the structure of the double helix provided a hint as to how DNA

divides and makes copies of itself. This model suggests that the two strands of the
double helix separate during replication, and each strand serves as a template from
which the new complementary strand is copied. What was not clear was how the
replication took place. There were three models suggested: conservative, semi-
conservative, and dispersive (see Figure 1).

Figure 1. The three suggested models of DNA replication. Grey indicates the original DNA strands, and blue
indicates newly synthesized DNA

In conservative replication, the parental DNA remains together, and the newly formed
daughter strands are together. The semi-conservative method suggests that each of
the two parental DNA strands act as a template for new DNA to be synthesized; after
replication, each double-stranded DNA includes one parental or “old” strand and one
“new” strand. In the dispersive model, both copies of DNA have double-stranded
segments of parental DNA and newly synthesized DNA interspersed.

Meselson and Stahl were interested in understanding how DNA replicates. They
grew E. coli for several generations in a medium containing a “heavy” isotope of
nitrogen (15N) that gets incorporated into nitrogenous bases, and eventually into the
DNA (Figure 2).
Figure 2. Meselson and Stahl experimented with E. coli grown first in heavy nitrogen ( 15N) then in 14N. DNA
grown in 15N (red band) is heavier than DNA grown in 14N (orange band), and sediments to a lower level in
cesium chloride solution in an ultracentrifuge. When DNA grown in 15N is switched to media containing 14N, after
one round of cell division the DNA sediments halfway between the 15N and 14N levels, indicating that it now
contains fifty percent 14N. In subsequent cell divisions, an increasing amount of DNA contains 14N only. This
data supports the semi-conservative replication model. (credit: modification of work by Mariana Ruiz Villareal)
The E. coli culture was then shifted into medium containing 14N and allowed to grow for
one generation. The cells were harvested and the DNA was isolated. The DNA was
centrifuged at high speeds in an ultracentrifuge. Some cells were allowed to grow for
one more life cycle in 14N and spun again. During the density gradient centrifugation, the
DNA is loaded into a gradient (typically a salt such as cesium chloride or sucrose) and
spun at high speeds of 50,000 to 60,000 rpm. Under these circumstances, the DNA will
form a band according to its density in the gradient. DNA grown in 15N will band at a
higher density position than that grown in 14N. Meselson and Stahl noted that after one
generation of growth in 14N after they had been shifted from 15N, the single band
observed was intermediate in position in between DNA of cells grown exclusively in 15N
and 14N. This suggested either a semi-conservative or dispersive mode of replication.
The DNA harvested from cells grown for two generations in 14N formed two bands: one
DNA band was at the intermediate position between 15N and 14N, and the other
corresponded to the band of 14N DNA. These results could only be explained if DNA
replicates in a semi-conservative manner. Therefore, the other two modes were ruled
out.

During DNA replication, each of the two strands that make up the double helix serves as
a template from which new strands are copied. The new strand will be complementary
to the parental or “old” strand. When two daughter DNA copies are formed, they have
the same sequence and are divided equally into the two daughter cells.

The model for DNA replication suggests that the two strands of the double helix separate during
replication, and each strand serves as a template from which the new complementary strand is
copied. In conservative replication, the parental DNA is conserved, and the daughter DNA is
newly synthesized. The semi-conservative method suggests that each of the two parental DNA
strands acts as template for new DNA to be synthesized; after replication, each double-stranded
DNA includes one parental or “old” strand and one “new” strand. The dispersive mode
suggested that the two copies of the DNA would have segments of parental DNA and newly
synthesized DNA. Experimental evidence showed DNA replication is semi-conservative.

The process of DNA replication is catalyzed by a type of enzyme called DNA

polymerase (poly meaning many, mer meaning pieces, and –ase meaning enzyme; so
an enzyme that attaches many pieces of DNA). During replication, the two DNA strands
separate at multiple points along the length of the chromosome. These locations are
called origins of replication because replication begins at these points. Observe
Figure 1: the double helix of the original DNA molecule separates (blue) and new
strands are made to match the separated strands. The result will be two DNA
molecules, each containing an old and a new strand. Therefore, DNA replication is
called semiconservative. The term semiconservative refers to the fact that half of the
original molecule (one of the two strands in the double helix) is “conserved” in the new
molecule. The original strand is referred to as the template strand because it provides
the information, or template, for the newly synthesized strand.
Figure 1. By Madprime(wikipedia) (DNA replication split horizontal) CC BY-SA 2.0

Figure 2. Primer and Template

DNA polymerase needs an “anchor” to start adding nucleotides: a short sequence of

DNA or RNA that is complementary to the template strand will work to provide a free 3′
end. This sequence is called a primer (Figure 2).
How does DNA polymerase know in what order to add nucleotides? Specific base
pairing in DNA is the key to copying the DNA: if you know the sequence of one strand,
you can use base pairing rules to build the other strand. Bases form pairs (base pairs)
in a very specific way.

Figure 3. DNA chemical structure. Modification of DNA chemical structure by Madeleine Price Ball; CC-BY-
SA-2.0

Now that you understand the basics of DNA replication, we can add a bit of
complexity. The two strands of DNA have to be temporarily separated from each other;
this job is done by a special enzyme, helicase, that helps unwind and separate the
DNA helices (Figure 4). Another issue is that the DNA polymerase only works in one
direction along the strand (5′ to 3′), but the double-stranded DNA has two strands
oriented in opposite directions. This problem is solved by synthesizing the two strands
slightly differently: one new strand grows continuously, the other in bits and pieces. The
leading strand grows continuously and the lagging strand is put together with short
pieces called Okazaki fragments. These fragments are connected and linked together
by an enzyme called DNA ligase. Short fragments of RNA are used as primers for the
DNA polymerase.
Figure 4. By Mariana Ruiz (DNA replication) Public Domain

Replication in eukaryotes starts at multiple origins of replication. A primer is required to initiate

synthesis, which is then extended by DNA polymerase as it adds nucleotides one by one to the
growing chain. The leading strand is synthesized continuously, whereas the lagging strand is
synthesized in short stretches called Okazaki fragments. The RNA primers are replaced with
DNA nucleotides; the DNA remains one continuous strand by linking the DNA fragments with
DNA ligase.