DNA replication is the production of new strands of DNA with base sequences identical

to existing strands

DNA replication is required for 2 biological processes

1. Reproduction: offspring need copies of the base sequence of their parents, so
parents must replicate their DNA to reproduce
2. Growth and tissue replacement in multicellular organisms: each cell needs a full set of
base sequences ∴ before a cell divides it must replicate all of its DNA. Cell division is
needed for growth and to replace cells in tissues where they have been lost.

This works through complementary base pairing. In new strands using this, strands
must be identical.

1. Each of the nitrogenous bases can only pair with its partner (A=T and G=C) this
is called complementary base pairing.

2.

The two new strands formed will be identical to the original strand.
Key: Blue: original, Green: new

3. Each new strand contains one original and one new strand, therefore DNA
Replication is said to be a Semi- Conservative Process.

The old strand unwinds, and then the old strand is used as a template. The new strand
is then made up against it. This keeps unwinding until they split. Each copy has one
blue, and green bit.

Semiconservative: Unwind blue, make green against it, and then the blue totally
unwinds. This makes two strips of DNA

Half of the old strand, half the new = DNA.

Out of the three originally proposed models (Semi-Conservative, Conservative,

Dispersive), semi-conservative was shown by Menellssohn and Stahl to be biologically
significant. This was worked out through showing that reproduced DNA has mixed
content (half-half)

2 Enzymes
•
Helicase
• The ‘ase’ ending indicates it is an enzyme
• This family of proteins varies but are often formed from multiple polypeptides and
doughnut-in-shape
 • Unwinds the DNA Helix (breaks it apart)
• Separates the two polynucleotide strands by breaking the hydrogen bonds
between complementary base pairs
• ATP is needed by helicase to both move along the DNA molecule and to break
the hydrogen bonds
• The two separated strands become parent/template strands for the replication
process.

DNA Polymerase
• The ‘ase’ ending indicates it is an enzyme
• This protein family consists of multiple polypeptides sub-units
• This is DNA polymerase from a human.
• The polymerisation reaction is a condensation reaction
• Free nucleotides are deoxynucleoside triphosphates
• The extra phosphate groups carry energy which is used for formation of covalent
bonds
• Present on both sides during replication

Artificial DNA replication - Polymerase Chain Reaction (PCR)

•

• Typically used to copy a segment of DNA – not a whole genome

• NEED: Gene of Interest on Double-Stranded DNA, Tag Polymerase,
Deoxyribonucleotides, DNA Primers, thermal cyclers, free DNA nucleotides
• Used to amplify small samples of DNA - Into billions of copies
• Typically 20 nucleotides long, to perform additional cycles
• Last several hours, 20-35 cycles
• With each cycle, the number of segments doubles
• IN CONJUNCTION with using them for DNA profiling, recombination, species
identification, or other research.

The Polymerase Chain Reaction (PCR): Animation

PCR is a way of producing large quantities of a specific target sequence of DNA. It is

useful when only a tiny amount of DNA is available for testing, e.g. crime scene
samples of blood, semen, tissue, hair, etc.

PCR occurs in a thermal cycler and involves a repeat procedure of 3 steps:

1. Denaturation: The DNA sample is heated to separate it into two strands
2. Annealing: DNA primers attach to opposite ends of the target sequence
3. Elongation: A heat-tolerant DNA polymerase (Taq) copies the strands

• One cycle of PCR yields two double stranded identical copies of the DNA
sequence
• A standard reaction of 30 cycles would yield 1,073,741,826 copies of DNA (230)
DNA Profiling
Compares sections of DNA between individuals in order to determine paternity or
relationships, as evidence in criminal cases or to identify species.

Through gel electrophoresis, fragments of DNA are moved through an electric field and
separated based on their size.

1. DNA samples are taken and amplified with PCR.

2. Restriction enzymes cut DNA into fragments at specific base sequences in
each sample.
3. A fluorescent marker binds to a triplet in the DNA fragments, so that results can
be seen.
4. Samples are added to a gel electrophoresis chamber. Electric current is passed
through, pushing the fragments along.
5. Heavier fragments (higher molecular weight) stay closer to the origin and
 smaller fragments go further.
6. A banding pattern shows up for each DNA sample and can be compared.

DNA profiling (electrophoresis) in paternity and forensic investigations

DNA is often left behind at a crime scene. It is present in all kinds of evidence, including
blood, hair, skin, saliva, and semen.
DNA can also be used in paternity investigations.
In 1986 forensic DNA analysis was first used. Originally known as "DNA fingerprinting,"
this type of analysis is now called "DNA profiling" or "DNA testing" to distinguish it from
traditional skin fingerprinting.
DNA samples are needed from the mother, (potential) father and child in question.
Reasons for investigations include:
• Inheritance of property, savings etc.
• Father is required to pay maintenance to support his biological child
It is easier to exclude a suspect than to convict someone based on a DNA match. The
FBI estimates that one-third of initial rape suspects are excluded because DNA samples
fail to match.
Forensic investigators take many precautions to prevent mistakes, but human error can
never be eliminated.
^To convict someone, mus thave more than 2 bands, must have STRONG
similarities.
1.

Ident
ify the smallest DNA fragment.

I. II. III. IV.

2. State the number of bands that would

appear in the ‘standard’ lane.

2 3 4 5 6

3. Identify the child which is most likely to be from the mother’s previous marriage.

1 2 3 4
In paternity, must have at least 50% match !