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DNA Replication- Maintaining
Genetic Information
 DNA must be copied, or replicated, so
that the information it holds can
maintained and passed to future cell
generations, even as that information is
accessed to guide the manufacture of
proteins.
 DNA Replication is the process by
which a double-stranded DNA molecule
is copied to produce two identical DNA
molecules.
DNA Replication is
Semiconservative
 The route to replication of DNA is called
semiconservative because each new
DNA double helix conserves half of the
original.
 Matthew Meselson & Franklin Stahl
(1957) – demonstrated the
semiconservative mechanism of DNA
replication with a series of “density shift”
experiments.
Steps of DNA Replication
 A site where DNA is locally opened,
resembling a fork, is called a replication
fork.
 DNA replication begins when an
unwinding protein called helicase
breaks the hydrogen bonds the connect
a base pair.
 Binding proteins hold / stabilize the two
single strands apart.
 Another enzyme, primase, the attracts
complementary RNA nucleotides to
build a short piece of RNA, called an
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RNA primer, at the start of each segment
of DNA to be replicated.
 The RNA primer is required because the
major replication enzyme, DNA
POLYMERASE (DNAP), can only add
bases to an existing nucleic acid strand.
o NOTE: A polymerase is an
enzyme that builds a polymer,
which is a chain of chemical
building blocks.
 Next, the RNA primer attracts DNAP,
which brings in DNA nucleotides
complementary to the exposed bases on
the parental strand that serves as a
mold, or template.
 The base-pairing rules ensure that each
base on the parental strand pulls in its
complement.
 Leading strand is the continuous strand
which synthesis continuously in 5’ to 3’
direction.
 Lagging strand is the discontinuous
strand that produces Okazaki fragments
on the 5’ to 3’ template.
 Growing fork procced in one direction
due to one strand which replication is
discontinuous.
 Next, an enzyme called a ligase seals
the sugar-phosphate backbones of the
pieces, building the new strand.
o Ligase comes from a Latin
word meaning “to tie”.
o These pieces, up to 150
nucleotides long, are called
Okazaki fragments.
 DNA polymerase also “proofreads” as
it goes, excising mismatched bases and
inserting correct ones.
 Yet, another enzyme, called an
annealing helicase, rewinds any
sections of DNA molecule that remain
unwound.
 Finally, ligases seal the entire sugar-
phosphate backbone.
 A human chromosome replicates
simultaneously at hundreds of points are
called replication bubbles.
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today can sequence an entire human

genome in a day.
 His method is still used to sequence
individual genes or to heck the accuracy
of a selected sequences part of a
genome.
 The ability to sequence millions of small
DNA pieces at once is the basis of newer
Polymerase Chain Reaction methods, called “next-generation
sequencing”, that can be handle much
 Researchers use DNA replication larger DNA molecules much faster.
conducted outside cells in biotechnology
called DNA amplification.
 The first and best-known DNA
amplification techniques is the
Polymerase Chain Reaction (PCR) –
which uses DNA polymerase to rapidly
replicate a specific DNA sequence in a
test tube.
 PCR is useful in forensic investigation to
amplify small DNA.
 STEPS IN AMPLIFYING DNA USING
PCR:
1. Select target sequence in
virus.
2. Primers
3. Copies of free nucleotides
4. Heat-resistant polymerase –
Taq1
5. Raised Temperature in target
sequence
6. Then lowered temp. to
separate two strands
7. Primers hybridize due to
base complementarity
8. DNA fills in
9. Repeat process many ties
 After 30 cycles, PCR yields more than
10 billion copies of the target sequence
Sequencing DNA
 Frederick Sanger – a British biochemist
and two-time Nobel Prize winner,
invented a way to determine the base
sequence of a small piece of DNA.
 “Sanger sequencing” remains the
conceptual basis for techniques that

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