U1 Gel electrophoresis is used to separate proteins or fragments of DNA according to size.

Gel electrophoresis: separating charged molecules in an electric field, according to their size and charge
Purpose: differentiate DNAs, RNAs, proteins based on structural and chemical differences
Ingredients
● Agar gel - jelly substance from algae, mixed with nutrients to serve as growth medium for
microorganisms
● Electrodes

Procedure
1. DNA samples amplified with PCR & restriction enzymes cut DNA into fragments
2. Insert DNA fragments into gel electrophoresis chamber
3. Run electric current through gel with (-) side on top
a. DNA phosphate group - negatively charged (all DNA molecules carry negative charges)
→ shorter samples will travel further in the gel (the gel has a web-like
structure)
→ more (-) samples will travel further in the gel
⇒ banding pattern shown up
 ● The gel used in gel electrophoresis consists of a mesh of filaments that resists the movement of
molecules in a sample
○ DNA molecules from eukaryotes are too long to move through the gel →
should be broken up to smaller fragments

U2 PCR can be used to amplify small amounts of DNA.

Polymerase chain reaction (PCR):
● Purpose: copying a small segment of DNA
● Ingredients
○ Target DNA strand
○ DNA primer
○ DNA polymerase
○ Nucleotides

1. DNA denaturation: spreads double-stranded DNA

a. Heat DNA at 94℃ for ~30 seconds
2. DNA primer: add primers that correspond to beginnings of target sequence
a. The primer binds to the start of the desired sequence and binds by complementary base
pairing
3. Annealing: cool temperature so that primers attach to target sequence
4. Add DNA polymerase: uses nucleotides to begin DNA replication

U3 DNA profiling involves comparison of DNA.

Modern use of PCR
● DNA amplification of extinct species
● Forensic investigation involving DNA
● Prenatal diagnosis for genetic disorders

DNA profiling
● A sample of DNA obtained
● Sequences of DNA selected and copied by PCR
● Copied DNA split into fragments using restriction endonucleases
● Fragments separated using gel electrophoresis
● Pattern of bands that is the same with DNA taken from one individual produced
○ This is the individual’s DNA profile
○ Human DNA has regions of short tandem repeats
○ Every individual has different number of repeats: Genomic fingerprint
● Profiles of different individuals compared

A1 Use of DNA profiling in paternity and forensic investigations.

Forensic investigations
● DNA profile of material from the crime scene is compared with the DNA profile of a sample DNA taken
from the suspect or the victim
Paternity investigations
● DNA profiles of the mother, the child, and the man are needed
● Patterns of bands compared
● If any band in the child’s profile does not occur in the profile of the mother or man, another person
must be the father.

U4 Genetic modification is carried out by gene transfer between species.

Genetic modification: transfer of genes from one species to another
● Possible because the genetic code is universal

A2 Gene transfer to bacteria using plasmids makes use of restriction endonucleases and DNA ligase.
● Plasmid: small, circular DNA molecules with relatively small numbers of genes
○ Usually contain genes that are beneficial but to essential to the bacterium
● Restriction enzymes (endonucleases): enzymes that
cut DNA molecules at specific base sequences
○ Different restriction enzymes cut DNA at
different DNA sequences called restriction sites
○ Can be used to cut open plasmids / cut out
desired genes from larger DNA molecules
○ Sticky ends created by a particular restriction
enzyme have complementary base sequences
so can be used to link together pieces of DNA
through hydrogen bonding between the bases
● DNA ligase: enzyme that joins DNA molecules together by making sugar-phosphate bonds between
nucleotides
○ DNA ligase seal the nicks in the sugar-phosphate backbone where the desired gene has been
inserted into a plasmid

● It is easier to obtain messenger RNA transcripts of genes than the genes themselves
● reverse transcriptase: enzyme that can create DNA strands based on the RNA template
● cDNA (complementary DNA): DNA copies of RNA molecules made by reverse transcriptase
Ethics of genetic modifications
Benefits
● Specific traits can be added or erased from organisms to benefit humankind
● ex. GM crop - golden rice: rice that contain beta-carotene gene that can be processed as vitamin A in
humans
Risks
● genetic modification can lead to mutations in certain genes, causing health problems
● Pest resistance gene in crops can spread to non-target wild grass, creating super weeds
● Unwanted genes can be spliced to create “designer babies”

A3 Assessment of the potential risks and benefits associated with genetic modification of crops.
Benefits and risks of GM crops
Benefits
● Environmental benefits
○ Pest-resistant crops can be produced - less insecticide has to be sprayed on the crop
○ Use of GM crop reduces the need for plowing and spraying crops - less fuel needed
○ Shelf-life of fruit and vegetables improved - reduces wastage and area of crops that have to
be grown
● Health benefits
○ Nutritional value of crops improved (ex. Golden rice)
○ Crops lacking allergens or toxins naturally present in them can be produced
○ GM crops that produce edible vaccines can be produced - by eating the crop, a person would
be vaccinated against a disease
● Agricultural benefits
○ Crops resistant to drought, cold, and salinity can be produced - increase total yields
○ Can make the crop herbicide resistant - the growing crop not killed when herbicide is
sprayed
■ Less weed competition → higher crop yields
○ Crops that are resistant to diseases caused by viruses produced

Risks
● Health risks
○ Proteins produced by transcription and translation of transferred genes could be toxic or
cause allergic reactions
○ Antibiotic resistance genes in GM crops can spread to pathogenic bacteria
○ Transferred genes can mutate and cause unexpected problems
■ ex. emergence of new allergens
● Environmental risks
○ Non-target organisms could be affected by toxins intended to control pests in GM crop plants
○ Herbicide resistant genes could spread to wild plants , turning them to uncontrollable
superweeds
○ Biodiversity can be reduced as many farmers use the same varieties of GM crops
● Agricultural risks
 ○ Widespread use of GM crops containing a toxin that kills insect pests will lead to the spread
of resistance to the tosin in the pests that were the initial problem
■ Spread of secondary pests resistant to the toxin
○ Farmers are prohibited by law from saving and re-sowing GM seed from crops they have
grown
■ Strains adapted to local conditions cannot be developed

U5 Clones are groups of genetically identical organisms, derived from a single original parent cell.
U8 Methods have been developed for cloning adult animals using differentiated cells.
A4 Production of cloned embryos produced by somatic-cell nuclear transfer.
● Cloning: production of genetically identical organisms
● Clone: a group of genetically identical organisms

● Adult somatic cell nucleus placed in enucleated egg cells to produce embryos that have the same
DNA as donor cell
● Clones have the same nuclear DNA as the donor, same mitochondrial DNA as surrogate mother

Procedure
1. Adult somatic cells taken from the udder of a Finn Dorset → grown in
laboratory in medium with low nutrients → this made genes in the cell
inactive, pattern of differentiation lost
2. Unfertilized eggs taken from the ovaries of a Scottish Blackface ewe →
nuclei removed → electric pulse used to fuse the somatic cultured cells
and the egg cell → fused cell developed into a embryo
3. The embryo injected into surrogate mothers

U6 Many plant species and some animal species have natural methods of cloning.
● Garlic bulb
○ A single garlic bulb grow leaves → leaves produce enough food by
photosynthesis to grow a group of bulbs
 ● Female aphids can give birth to offspring that have been produced entirely from diploid egg cells
that were produced by mitosis

U7 Animals can be cloned at the embryo stage by breaking up the embryo into more than one group of
cells.
● Individual cells can be separated from the embryo while they are still pluripotent and transplanted
into surrogate mothers
● Embryonic stem cells (ES sells) are undifferentiated and thus can reproduce & develop into all
varieties