DNA

Replication
D1.1
Oxford pg. 105 - 122
Pearson pg. 575-582
Guiding Questions

 How is new DNA produced  How has knowledge of DNA

replication enabled
applications in biotechnology
 CRISPR

 Technology for manipulating  Leads to ethical questions

DNA sequences and implications
 Can be used to cure disease o Altering embryos to become
rather than just treating star athletes
them o High IQ levels
o Huntington's disease o "perfect" physical attributes
o Duchene muscular dystrophy o High immune system levels
o Cystic Fibrosis  What does this look like for
evolution?
DNA analysis results

 Family ancestry
 Forensic investigations
 Inheritance of medical conditions
 Sequencing DNA of a million-year-old woolly mammoth
 Extracted cartilage from a 125-million-year-old Caudipteryx
dinosaur, not sequenced yet
D1.1.1 The role of DNA replication

DNA replication as production

of exact copies of DNA with
Identical base sequences
 Students should
know
that DNA replication is required for
reproduction and for growth and tissue
replacement in multicellular organisms
DNA

 Polymer made of monomers  Provides the code for all life

(nucleotides) forms
 Sugar (deoxyribose),  Must be duplicated
phosphate group, and o Reproduction
nitrogenous base (A/T, C/G)
o Growth
o Tissue replacement
o Copies must be exact
 DNA types

Mitochondrial Nuclear
 Only inherited from the
maternal lineage  Inherited from both
parents
 Circular
 Linear
 In cell mitochondria
 In cell nucleus
 Majority of DNA, is the
genetic directions
D1.1.2 Semi-conservative Replication

Semi-conservative nature of
DNA replication and role of
complementary base pairing
 Students should
know
How these processes allow a high degree of
accuracy in copying base sequences
 Allows for a high Semi-
degree of accuracy
in copying base
sequences and
conservati
provides stability
when passing on the ve
genetic code
replication
DNA replication ensures two identical copies of DNA are produced
from an original strand. Based on complementary pairs of
nucleotides (base pairs A=T, C=G)
 DNA
replication
is Semi-
Conservativ
e

EACH STRAND IN THE

DOUBLE HELIX ACTS AS A
TEMPLATE FOR THE
SYNTHESIS OF A NEW,
COMPLEMENTARY STRAND
D1.1.3 Complementary base pairing

Role of helicase and DNA

polymerase in DNA
replication
 Students should
know
limit to the role of helicase in unwinding and
breaking hydrogen bonds between DNA strands
and the general role of DNA polymerase
 Enzyme
Separates double-
stranded DNA into
Helicase single strands
Breaks hydrogen
bonds between base
pairs
 Enzyme that adds
nucleotides one at a
DNA Polymerase time
Grows the DNA
chain
 D1.1.4 Amplifying and Separating DNA

Polymerase Chain reaction and gel

electrophoresis as tools for amplifying
and separating DNA
 Students should
know
The use of primers, temperature changes, and
Taq polymerase in the polymerase chain
reaction (PCR) and the basis of separation of
DNA fragments in gel electrophoresis
 Sometimes called "molecular
photocopying," the polymerase chain
reaction (PCR) is a fast and inexpensive
technique used to "amplify" - copy -
small segments of DNA. Because

Polymeras
significant amounts of a sample of DNA
are necessary for molecular and genetic
analyses, studies of isolated pieces of

e Chain
DNA are nearly impossible without PCR
amplification.
Once amplified, the DNA produced by

Reaction PCR can be used in many different

laboratory procedures. For example,
most mapping techniques in the Human

(PCR) Genome Project (HGP) relied on PCR.

PCR is also valuable in a number of

laboratory and clinical techniques,
including DNA fingerprinting, detection
of bacteria or viruses (particularly
AIDS), and diagnosis of genetic
disorders.
Polymerase Chain
Reaction (PCR)
How does PCR Work?

To amplify a segment of DNA using PCR, the sample is first heated so the DNA denatures,
or separates into two pieces of single-stranded DNA. Next, an enzyme called "Taq
polymerase" synthesizes - builds - two new strands of DNA, using the original strands as
templates. This process results in the duplication of the original DNA, with each of the
new molecules containing one old and one new strand of DNA. Then each of these
strands can be used to create two new copies, and so on, and so on. The cycle of
denaturing and synthesizing new DNA is repeated as many as 30 or 40 times, leading to
more than one billion exact copies of the original DNA segment.

The entire cycling process of PCR is automated and can be completed in just a few hours.
It is directed by a machine called a thermocycler, which is programmed to alter the
temperature of the reaction every few minutes to allow DNA denaturing and synthesis.
 PCR Vocab

 Primers – single-stranded, short polymers, complementary to the nucleotides at

one end of the DNA strand, provide a starting point for DNA synthesis
 Taq polymerase – from a bacterium in hotsprings, can withstand high
temperatures
 Free nucleotides – bases uses to create new DNA strand
 Denaturation – step 1 – mixture is heated to 92 C – 98 C, hydrogen bonds broken
 Annealing – step 2 – mixture cooled to 50 C – 65 C, primers bind with nucleotides
at ends of target sequences
 Elongation – step 3 – mixture at 70 C – 80 C, Taq polymerase catalyses the
building by extending the primers
All About PCR - Beta - click the link and follow along
Virtual PCR Simulat
or

Try this lab

Gel Electrophoresis
Try this lab
D1.1.5 Applications of Amplifying and
Separating DNA

Applications of polymerase
chain reactions and
electrophoresis
 Students should
know
Appreciate the broad range of applications,
including DNA profiling for paternity and
forensic investigation
 Nature of science
Reliability is enhanced by increasing the
number of measurements in an experiment or
test. In DNA profiling, increasing the number of
markers used reduces the probability of a false
match
DNA Profiling

 Matching an unknown DNA sample with a known sample

 After separation via gel electrophoresis, patterns of bands are compared. identical
pattern = same individual DNA, similar pattern = related
 Used for paternity tests
 Used at crime scenes and criminal cases – when little DNA is collected, PCR is done to
make more DNA available for testing
 Used in studies of ecosystems – DNA of non-humans are also studied, helps
understand animal relationships and evolution
 Guiding Questions

 How is new DNA produced

o DNA replication is essential to produce new cells, both in unicellular and
multicellular organisms
o Semi-conservative replication results in every new copy of DNA
possessing one strand from the parent DNA and one new strand
o Complementary base pairing in the DNA replication process ensures a
high degree of accuracy
o Helicase is required to unwind and break hydrogen bonds so that the
process of DNA replication can occur
o DNA polymerase controls the addition of free nucleotides to the
developing DNA strand
 Guiding Questions

 How has knowledge of DNA replication enabled applications in biotechnology

o Polymerase chain reactions (PCR) are used to amplify DNA segments so that valid
analyses can be carried out
o A thermocycler, Taq polymerase, primers, and free nucleotides are necessary for
the process of PCR
o Gel electrophoresis uses an electrical field and agarose gel to separate segments of
DNA
o PCR and gel electrophoresis are often used together in attempts to solve crimes
where small amounts of DNA have been recovered
o Gel electrophoresis is also used in the study of ecosystems
o Paternity questions can often be solved using gel electrophoresis
Question Review
210 amino acids = 630 nucleotides (3 bases = 1 amino acid)
MRNA and DNA have covalent bonds between adjacent nucleotides
Why is TAQ DNA polymerase used – does not denature at high
temps
Amino acids joined to polypeptides – in ribosomes
Replication, transcription, and translation – all have complementary
base pairing
Twins show different methylation patterns due to – different
environment/illness/disease/diet
 Sons have 50% of having hemophilia
Daughters have 0% having it/ 50% being a
carrier
Blood clotting process

 blood clotting seals cuts in the skin;

 clotting factors are released (from platelets);
 thrombin is activated (enzyme of blood coagulation)
 a cascade reaction occurs (with thrombin);
 (thrombin causes) fibrinogen is converted to fibrin (protein)
 fibrin forms a clot/blocks the cut/prevents blood from being lost;
 Consequences of Hemophilia

 if a person does not have enough clotting factors/hemophilia, the

clot will not form;
 pathogens can enter the body more easily;
 (in hemophiliacs) blood will be lost from a cut which affects blood
pressure/bleeding to death;
 loss of blood affects amount of hemoglobin/O2 carried around the
body;
 reference to lifestyle / menstrual/birth problems
 Outline the stages in the production of
mRNA by transcription
a. DNA is unwound/strands are separated «by RNA polymerase»
b. new nucleotides attached to template strand «by RNA
polymerase»
c. complementary base pairing/base pairing with an example /
adenine with thymine/ uracil with adenine/ cytosine with guanine/
guanine with cytosine
d. mRNA detaches from template
e. DNA rewinds
 a. decided to combine what
was known about chemical
content of DNA with

How did information from X-ray

diffraction studies
b. built scale models of
model-making components of DNA
c. then attempted to fit them

help Watson together in a way that agreed

with the data «from separate
sources»

and Crick? d. made several arrangements

of scale model until found best
one that fitted all the data
 a. associated with
Distinguish
«histone» proteins in
eukaryotes but not
between
prokaryotes chromosomes
b. is linear in
eukaryotes but of eukaryotic
circular in prokaryotes
c. in cytoplasm in
and
prokaryotes, but
within nucleus in
prokaryotic
eukaryotes. cells
 (nucleosomes can) promote AND inhibit
transcription of genes/expression of genes;

Outline how (nucleosomes can) prevent transcription

by (tight)
nucleosome condensation/supercoiling/packing of DNA;
(nucleosomes can) allow/prevent binding
s affect the of RNA polymerase/transcription factors;
tagging/acetylation/methylation of
transcriptio nucleosomes/histones can promote/inhibit

n of DNA transcription;
movement of histones/nucleosomes
(along DNA) can affect which genes are
transcribed;
 Explain the role of lactose in the
expression of the gene for lactase
production

- lactose binds to repressor protein;

- repressor protein (with lactose bound) cannot block/bind to the
promoter/Y;
- RNA polymerase/X binds to the promoter/transcribes the gene;
- lactase produced (if lactose present)/lactase production
inhibited if lactose absent;