Heredity
Introduction to Reproduction
Mechanisms of Reproduction
What is Reproduction
reproduction is the production of offspring
it is necessary for the continuation of life
what are ways can organism can reproduce
sexual reproduction
2 parents are involved, one male and one female —> each provide half of the genetic information for the new offspring in the form of a sex cell (sperm and egg)
similar characteristics to parents but NOT identical
Asexual reproduction
1 parent and no fertilisation
genetically identical to parent - they are clones
Reproduction in Different Organisms
Animals
Sexual reproduction
male sperm and female egg fuse together during fertilisation to form a new organism
what are the different types of fertilisation
external
fertilisation outside of the female body
both the male and female release their sex cells directly into their surrounding environment
fish and amphibians fertilise externally
advantages:
large number of offspring can be produced because of the huge number of sex cells that can be released
can allow fertilisation to occur without the parents meeting
disadvantages:
environmental conditions mean that the chances of the sex cells surviving are reduced
wasted resource as many of the sex cells released do not get the chance to fertilise
no guarantee any offspring will survive
internal
fertilisation within the female’s reproductive system
usually land based - mammals, reptiles and birds fertilise internally
advantages:
offspring have higher chances of survival because the environment inside of the female body is more contained and protects sex cells
organism can save on resources and produce less sex cells
allows the animal to choose its mate
disadvantage:
only a few offspring can be produced at once
parents must physically meet
Plants
sexual reproduction
pollination
insects, birds or wind transport pollen (male sex cells) from one flower to another deep inside the plant are ovules (female plant sex cells) after fertilisation the female part of the flower develops into a fruit and the ovules turn into the seeds
asexual reproduction
runners:
side branches with little clumps of leaves and roots called plantlets distributed along them. They grow along the ground and the roots dig down and establish the plantlets as individual plants
bulbs:
from the bulbs little buds form and grown form new plants
cuttings
branch from the parent plant is cut off and planted into the ground where it will grow roots and establish itself as a new plant
Fungi
Fungi are eukaryotic organisms that can be either unicellular or multi-cellular
can reproduce sexually or asexually
determination of which reproduction to use it based on the environment
good conditions —> reproduce asexually and spread quickly (budding or spores)
bad conditions —> reproduce sexually through spores as two spores from two different fungi fuse together to form a single new cell
sexually increases genetic variation and he species chances of survival
Bacteria
Bacteria is prokaryotic single celled organism
asexual reproduction - Binary fission
cell replicates its DNA
cell grows to about twice its original size
cytoplasm divides and a new cell membrane forms
Protist
Protist are eukaryotic single celled organism
asexual reproduction - binary fission
Evaluating Reproduction
Comparing Sexual and Asexual Reproduction
features | sexual reproduction | asexual reproduction |
number of parents | two | one |
genetics of offsprings | unique | identical |
type of cell division | meiosis | mitosis, binary division, vegetative propagation and budding |
advantages |
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|
disadvantages |
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organisms | animals, plants, fungi | plants, fungi, bacteria, protists |
Mammalian Reproduction
what is a mammal
warm-blooded
vertebrate (has a backbone)
lactates (able to produce milk)
have hair
mammalian reproduction
fertilisation refers to the fusion of the egg and sperm
internal fertilisation occurs within the female’s reproductive tract
implantation refers to the attachment of the fertilised egg to the uterine lining
pregnancy refers o the time the offspring spends within the female’s body
fertilisation
ovulation - ovary releases egg
egg travels through the fallopian tube (oviduct)
male releases semen (containing sperm) during sex
the egg and sperm fuse to create a zygote
zygote forms strong outer membrane to stop more sperm entering
implantation
the zygote travels down to the cervix where it buries deep into the lining of the uterus
as the embryo develops the amniotic sac, placenta and umbilical cord form
amniotic sac —> bag containing fluid which helps keep embryo at optical temperature and provides cushioning
placenta —> provides nutrients and removes waste
umbilical cord —> connects offspring to placenta where it allows substances to move between
role of hormones
hormones are the chemical messengers produced by the body, which travel in the blood to other cells where they have a specific effect
Oestrogen
made by the placenta
stimulates ovulation
aids blood flow to offspring
aids organ development
stimulates progesterone
progesterone
progesterone is made by the ovaries, and them by the placenta
stimulates thickening of uterus lining
aids placenta function and relaxes uterus
helps mother’s immune system tolerate infant
other hormones
relaxin
loosens uterine muscles during pregnancy
oxytocin
stimulates production of milk
triggers uterine contractions
Manipulation in Agriculture
Reproduction in Agriculture
What is Agriculture
Agriculture is the growth of crops and animals for human needs
provides us with food and clothing
Manipulating Reproduction in Agriculture
selective breeding refers to the creation of organisms with certain desirable characteristics
Natural Breeding
involves placing a male and female from the same species in a enclosed environment, and waiting for them to mate
this is the most simple and easy way to manipulate reproduction
Artificial Insemination
involves taking sperm form a male (who has desirable traits) and inserting it directly into the female’s reproductive tract
Artificial Pollination
involves taking pollen from one flower and inserting it directly into another flower using a small brush
cloning
involves creating a genetically identical copy of an organism using genetic engineering techniques
this is the most complex way to manipulate reproduction
Impacts of Manipulating Reproduction on Agriculture
advantages:
increased sales
increased production
improved food quality
increases resistance to certain pests and diseases
farmers save money
less environmental contamination
disadvantages:
reduced biodiversity (if used excessively)
as ‘undesirable’ traits are bred out, variation in a species decreases which means the population is more likely to suffer from changes in the environment
DNA and RNA
DNA and RNA
What does DNA do
a type of nucleic acid that is responsible for storing genetic information in cells
acts as an instruction manual: contains information needed for growth, survival and replication
eukaryotes
relatively large amount of DNA
DNA is bounded
forms linear chromosomes
chromosomes reside in the nucleus
prokaryotes
relatively small amount of DNA
DNA is unbounded
have a single, circular chromosome
chromosome reside in the cytoplasm
DNA structures
consists of two chains of building blocks that twist together form a double helix
consists of building blocks called nucleotides
Deoxyribose sugar
Phosphate group
Nitrogenous base
Guanine
Cytosine
Adenine
Thymine
each base has a complementary base pairing
A + T
C + G
this ensures the ‘rungs’ are all the same length
deoxyribonucleotide join together forming a long chain
two chains twist around each other to form a double helix structure
the deoxyribose sugars and phosphate groups are on the outside whereas the nitrogen bases make up the rungs on the inside
What is RNA
a type of nucleic acid that is responsible for interpreting genetic information (from DNA) into proteins
DOES NOT STORE GENETIC INFORMATION
helps to turn DNA into proteins

RNA Structure
consists of nucleotides which contain sugar, phosphate and nitrogen base
RNA is single stranded
there is a specific type of nucleotide in RNA called ribonucleotide
RNA does NOT have thymine they HAVE uracil
G + C
A + U

DNA Replication
as cells are constantly replicating, DNA has to as well
unravelling the DNA

the enzyme DNA helicase breaks the hydrogen bonds between the bases, ‘unzipping’ the double helix into two single strands
Building a new strand

semi- conservative: when DNA is produced, one of the strands in each new DNA molecule comes from the old DNA molecule
reduced the chance of copying a strand incorrectly
know how to draw an annotated diagram of a DNA replication or whatever
DNA polymerase helps bind free nucleotides in the cell nucleus to the single strand

Draw a labelled diagram demonstrating the difference in the form of DNA between prokaryotic and eukaryotic cells.

DNA is Prokaryotes and Eukaryotes
Prokaryotes
a cell is prokaryotic if it does NOT have a nucleus or any other membrane bounded organelles
DNA is found in the cytoplasm, and, it is small and circular
examples
bacteria and archaea
usually unicellular
earliest life forms
Eukaryotic
a cell is eukaryotic if it DOES have a nucleus and membrane bound organelles
DNA is found in the nucleus and is linear
has lots more DNA compared to prokaryotes
examples
animals and plants
multicellular (cell has organelles and can perform more complex and specialised functions)
DNA in Prokaryotes vs Eukaryotes
DNA features | Prokaryotic cells | Eukaryotic cells |
DNA shape | Circular | Linear |
DNA location | Cytoplasm | Nucleolus |
Relative amount of DNA | Small | Large |
DNA and Replication
DNA replication
Unravelling the DNA
DNA is made up of two polynucleotide stands held together by hydrogen bonds between complementary bases
The enzyme DNA helicase breaks the hydrogen bonds between the bases, ‘unzipping’ the double helix into two single strands

building a new strand
helicase is still acting on the strands
the exposed strands act as templates for a new strand

semi-conservative: when DNA is produced one of the strands in each new DNA molecule comes from the old DNA molecule
reduced the chance of copying a strand incorrectly
know how to draw an annotated diagram of a DNA replication
DNA polymerase helps bind free nucleotides in the cell nucleus to the single strand

Forming the DNA backbone
DNA polymerase stitches these newly joined nucleotides together so that the sugar-phosphate backbone is formed
initially, only the bases are held together by weak hydrogen bonds, the new strand is then reinforced by the sugar-phosphate backbone
in baby words:
Bases Pair First: The free nucleotide bases (A, T, C, and G) pair up with their complementary bases on the original DNA strand using weak hydrogen bonds. This forms the "steps" of the ladder.
DNA Polymerase Builds the Backbone: DNA polymerase connects the sugar of one nucleotide to the phosphate of the next nucleotide, forming the sugar-phosphate backbone. This backbone runs along the outside of the DNA strand and gives it strength.
two identical DNA molecules are produced!

Mitosis and Meiosis
cell replication is the process by which cells replicate their genetic material and divide to form new cells
Mitosis
is a type of cell division where 1 parent cell divides once to produce two identical daughter cells
DNA is located in the nucleus and is wrapped around in a protein, this is called a chromatin
parent cell:


usually there are lots of smaller pieces of chromatin’s rather than just one big one
half of the chromatins comes from the mother and the other half comes from the father
step 1 involves the replication of DNA / chromatin
the two identical chromatin pieces are still joined in the middle and they quickly shorten and thicken
when coiled up in the X shape it is called a chromosome

the chromosomes come in matching pairs, also known as homologous chromosomes
step 2: chromosomes line up individually
step 4: spindle fibres attach to centromeres

step 5: spindle fibres shorten, centromeres break and chromatids move apart
step 6: cytoplasm separates, cell and nuclear membranes from, chromosomes uncoil

identical daughter cells
name | definition |
DNA replication | DNA replicates |
Prophase | Chromatin shorten and thickens to form chromosomes |
Metaphase | Chromosomes line up individually |
Anaphase | Spindle fibres shorten, centromeres break and chromatids move apart |
Telophase | Cytoplasm separates, cell and nuclear membranes form, chromosomes uncoil |
why is mitosis important
it creates new body cells that are needed for growth, repair, and maintenance
important for the individual
allows us to heal injuries, grow and survive
important for the species
increases and organism’s chances of reproducing
Meiosis
is a type of cell division where 1 parent cell divides twice to produce 4 non-identical daughter cells

MEIOSIS I
step 1: DNA replication
step 2: chromatin shortens and thickens to form chromosomes
step 3: chromosomes line up in homologous pairs

step 4: crossing over occurs (homologous chromosomes swap sections)

step 5: spindle fibres attach, shorten, and homologous chromosomes break move apart

MEIOSIS II
step 6: chromosomes line up individually
step 7: spindle fibres attach to centromeres
step 8: spindle fibres shorten, centromeres break and chromatids move apart

step 9: cytoplasm separates, cell and nuclear membrane form, chromosomes uncoil
step 10: four non-identical daughter cells (gametes / sex cells)
why is meiosis important
allows for the continuity of a species because it means that organisms can produce offspring with new gene combinations
evolution (natural selection): a population that has variation is more likely to survive environmental changes

Cell Replication In Practice
Sources of Genetic Variation in Sexual Reproduction
Genetic Variation
is a term used to describe the differences between the genomes of individuals of the same species
genome - an organism’s complete set of DNA
species - group of organisms which can interbreed to produce live, fertile offspring
differences in the DNA of a group of similar organism
where does this variation come from?
Random Segregation and Crossing over (Meiosis)
independent assortment - pairs of homologous chromosomes arrange independently to each other,
as a result
random segregation - when the chromatids segregate the set of chromosomes in the daughter cells is random (different combinations = genetic variation)
crossing over also contributes to genetic variation because it changes the genetic composition of the chromosomes
Fertilisation
contributes to genetic variation because it allows for different gametes (containing different alleles) to combine
there are lots of unique organism in a species, which all have lots of unique gametes (due to meiosis). Any of these gametes can combine during fertilisation to form different zygotes
mutation
refers to a change in the base sequence of an organism’s DNA
a change in the DNA = different genes = different traits = different phenotype
DNA and Polypeptide Synthesis
Key terms in Genetics
Genome
is an organisms complete set of genetic material (DNA)
it includes all of the information the organism needs to grow and survive
gene
a section of genetic material (DNA), each gene codes for a different characteristic
the different sections of the genetic information control a particular feature of the organism
each gene acts as a template for a particular mRNA strand which is translated into a particular polypeptide which determines a particular characteristic
alleles
alleles are alternative forms of the same gene (that’s why people have different hair and eye colours)
alleles are found at the same place on the same chromosome (in different organism from the same species)
genotype
refers to an organism’s genetic make-up for a particular characteristic
phenotype
refers to an organism’s physical expression of a particular characteristic
it is determined why two main things
which genes are present (genotype)
environmental factors

Genotypes and Phenotypes
Genotype
refers to an organism’s genetic makeup for a particular characteristic
typically an individual’s genotype for a trait consists of two alleles, one inherited from their mother and one from their father
genotype is determined at fertilisation, when a sperm and egg fuse to form a zygote
Phenotype
refers to an organism’s physical expression of a particular characteristic
it is observable or expressed characteristics of an organism
determined by which alleles are present (the genotype) as well as environmental factors
Proteins
Structure
consists of building blocks called amino acids which are linked together via peptide bonds
amino acids are made of carbon, hydrogen, oxygen and nitrogen
proteins are the only biological molecules that contain nitrogen

Function
two classifications of protemone
structural proteins: maintain cell shape and make out connective tissues
collagen which increases the flexibility of skin and bone
enzymes: biological catalysts - living proteins which speed up chemical reactions occuring in cells. they act on specific substrates to either break it down or combine it
sucrase breaks sucrose down into glucose and fructose
DNA polymerase bonds nucleotides together during DNA replication
hormones: proteins which are secreted into the blood by endocrine cells (glands) - travel to target tissues, where they cause a change in activity
insulin which regulates blood glucose
immunity: antibodies are proteins involved in the immune response - antibodies react with antigens (foreign particles) to help remove them from the body
Testing for Proteins (Biuret Test)
add a few drops of sodium hydroxide to the sample (makes the solution alkaline which is required for the test to work)
add a few drops of copper sulfate
if it remains blue, protein is not present but if it turns purple then protein is present
Protein Synthesis
Introduction to Genes
gene: a portion of DNA that tells the cell how to make a specific protein
codon: each set of three codes for one particular amino acid in the polypeptide chain
the order of codons and the bases within them determine which amino acids are produced and in what order
special codons that say when to start and stop reading code
translating DNA is like code breaking, and the genetic code is the template
specific codon —> specific amino acid
different codons can code for the same amino acid
the genetic code if ‘degenerate’
protein synthesis
transcription
an mRNA copy of a gene is made using DNA as a template
takes place in the cell nucleus (1)
RNA polymerase attaches to the DNA at the desired gene and separates the strands to expose the nucleotides in that region (only the section with the gene is pulled apart, NOT the whole DNA strand) (2)
mRNA is made by complementary base pairing, not matching identical bases
RNA polymerase adds RNA nucleotides one at a time (3)
DNA strands rejoin so the bases are not left exposed

in prokaryotes this is where it is completed but in eukaryotes there is one more step
in eukaryotes, introns are spliced out of the strand and exons are stuck together to form the final mRNA strand
introns: don’t code for proteins
exons: expressed, code for proteins
mRNA molecules leave the nucleus and enter the cytoplasm through a nuclear pore to the cytoplasm where it attaches to a ribosome for translation
translation
nucleic acid is translated into protein. A polypeptide chain is formed according to the sequence of codons
mRNA attaches to the ribosome at a particular start codon
anticodon: a triplet bases that corresponds to a specific mRNA codon
the tRNA molecule with the correct anticodon pairs up with mRNA in the ribosome
each tRNA molecule is bound to a specific amino acid
a second tRNA molecule attaches to the next codon on the mRNA strand
the ribosome catalyses the formation of a peptide bond between the two amino acids
the first tRNA molecule moves away from the ribosome leaving its amino acid, these repeats for the other tRNA until the stop codon where the last tRNA is bound to the ribosome
when a stop codon is reached the mRNA, tRNA and ribosome separate, leaving a finished polypeptide chain

Genetic Variation
Inheritance
introduction to inheritance
inheritance explains how characteristics are passed from one generation to the next, based on the transfer of DNA
multiple alleles
alleles are different versions of the same gene, and are found at the same place on homologous chromosomes
dominant and recessive alleles
dominant allele is always expressed in the phenotype
the recessive allele is only expressed in the phenotype when there is no dominant allele
a heterozygous of non-pure-breeding organism has 2 different alleles for a particular gene (brown and blue eye allele)
homozygous or pure-breeding organism has 2 identical alleles for a particular gene (brown and brown eye allele) (blue and blue eye allele)
punnett squares
predict the possible genotypes and phenotypes of the offspring produced by a particular male and female


as every gamete has the dominant tall allele (T), 100% of the pea plants will be tall
autosomal and sex-linked inheritance
types of chromosomes
humans and 23 homologous (matching) pairs of chromosomes (46 in total)
each pair of homologous chromosomes codes for the same genes
classified into two categories based whether they code for sexual characteristics
sex chromosomes
female: 2 similar chromosomes (XX)
male: 2 dissimilar chromosomes (XY)
autosome
don’t code for any sexual characteristic - normal
autosomal inheritance
patterns in the expression of characteristics which are found on autosomes
dominant/recessive
co-dominant
incomplete
sex-linked inheritance
patterns in the expression of characteristics which are found on sex chromosomes (X and Y)
sex-linked inheritance is different to autosomal inheritance due to 2 facts
the y chromosome has fewer genes than the x chromosome
males only have one copy of the x chromosome
x-linked
transmission of genes which are found on the x chromosomes
recessive: affect men more than woman as there is a higher chance of a man inheriting one faulty x chromosome
dominant: affects gender equally
y-linked
transmission of genes which are found on the y chromosome
only passed from father to son: never seen in females

Codominance and incomplete
Codominance
refers to a situation in which both alleles are expressed in the phenotype
use different capital letters to represent codominant alleles in punnett squares

example: human blood type
blood type consists of A, B, AB, and O
a person’s blood type is determined by their combination of alleles
there are multiple alleles for blood type: by I^A, I^B and i
these alleles code for glycoproteins in the outer membrane of red blood cells

incomplete dominance
the dominant allele is only partially expressed - it does not completely control the phenotype of a heterozygous organism
red flower and white flower make pink flower = the red allele only partially masks the white allele resulting in the pink phenotype
Pedigrees
features of pedigrees
diagram which shows how traits are passed on over many generations
applications of pedigrees
identify patterns
determine genotypes
predict phenotypes
Reading Pedigrees
Examining Data
Allele Frequency
allele frequency refers to the relative proportion of a particular allele in a population
allele frequency = number of an allele in a population / total number of alleles in the population
you can work out allele frequencies by taking a sample of the population, as long as the sample is big enough for us to say that it is likely to be representative
allele frequencies can be calculated from information about alleles, or by looking at phenotypes so long as we know the relationship between genotype and phenotype.
The Hardy-Weinberg Model
describes and predicts alleles frequencies in a non-evolving population
states the alleles frequencies will stay the same across generations as long as no evolutionary processes occur
is used to calculate genotype frequencies in a population, based on given alleles frequencies
Predicting Speciation
there are several evolutionary processes which can cause populations to move away from the predictions of the hardy-Weinberg model, including
natural selection
preferential mating
migration
mutations
and random events
the influence of evolutionary processes means that the model can not always accurately predict allele and genotype frequencies in a population
if allele frequencies in a population are not following the hardy-Weinberg equilibrium we can assume that the speciation is occurring
speciation typically occurs when there is a barrier to gene flow
if the allele frequencies in two populations are different there is probably a barrier to gene flow
if we measure frequency data for enough alleles we many be able to work out of two populations have stopped interbreeding making them different species
Population Genetics
Population Genetics in Conservation
measuring extinction risk
location: Galapagos Island, what: heavy rains, impact: cactuses produced smaller seeds
the change seeds had knock-on effects for species feeding on the seeds, such as the common cactus finch
if the finch population has high genetic diversity
variation in observable traits exists (including beak size)
finches with smaller beaks can easily eat the smell seeds
finches with smaller beaks more likely to survive than finches with larger beaks
in finches population has low genetic diversity
less variation in genes contributing to beak size
lower number of finches with favourable small beaks
population could be significantly reduced or eradicated
populations with greater genetic diversity withstand environmental changes better
conservation scientists measure population genetics to calculate the risk of extinction
monitoring inbreeding
inbreeding refers to when closely related individuals breeding together (this occurs when individuals only have access to a small range of potential mates)
closely related individuals have similar ancestors, so are more likely to inherit the same unhealthy allele
unhealthy alleles are often recessive
inheriting two copies of the unhealthy allele led new born choughs to die soon after birth
the high death rate among young birds led the Scottish Chough population to fall rapidly.
inbreeding may occur if population size suddenly falls or if population is split into smaller groups
comparing the genomes of a random population sample can allow us to monitor inbreeding by:
allowing us to determine how closely related population members are
allowing us to measure the allele frequencies for unhealthy mutations
allele frequencies —> the relative proportion of a particular allele in a gene pool
conservation scientist look at population genetics to monitory inbreeding
investigating speciation
speciation refers to the formation of new species
species —> a group of organisms that interbreed to produced fertile offspring
cryptic species are groups of species originally through to be the same but are different
during speciation, populations stop inbreeding, preventing DNA from being shared between populations, creating a barrier to gene flow
alleles may be lost in one population and not the other
new alleles ay arise in one population and not the other
population genetics allows us to compare the allele frequencies within and between populations
if allele frequencies are becoming very different, it suggests speciation is occurring
conservation scientist look at population genetics to better understand ecosystems
Population Genetics in Disease Inheritance
disease affected by multiple genes
many genetic disorders are affected by more than one gene
polygenetic inheritance refers to when the inheritance of an observable trait is determined by many genes
genes showing polygenic inheritance often have environmental influences such as cardiovascular disease and some mood disorders
identifying the regions of DNA associate with a genetic disorder shows:
who is likely to be affected
what goes wrong in the cell to cause the disease
how we can treat the disease
Genome-Wide Association Studies
can be used to identify single nucleotide polymorphisms (SNPs) that affect you chance of inheriting a genetic disease
contribute to risk of getting a disease
are found near a region of DNA contributing to the risk of getting a disease
can be associated with a particular ethnicities of heritages
population stratification can be used to ensure we identify SNPs associated with disease rather than ethnicities
Population Genetics in Human Evolution
primate evolutionary relationships
humans are classified as Homo sapiens
we can base our ideas about the evolutionary relationships of different species on their observable characteristics as well as DNA evidence
DNA and Evolutionary Relationships
As humans share 99% of their DNA with chimpanzees (and only 98% with gorillas) they share ore recent common ancestor with and are more closely related to chimpanzees than to gorillas
there has been more time for the DNA of humans and gorillas to evolve differences
the more we collect and analyse DNA samples the more accurate our understanding of primate evolution is
on average, each base pair of primate DNA has a one in a billion chance of mutating per year
studying human migration
analysing DNA can inform us of evolutionary relationships
if we know the rate of DNA mutation we can work out when the last common ancestor of two species of populations lived
Inheritance Patterns in a Population
PCR and Gel Electrophoresis
PCR
what is PCR
Polymerase Chain Reaction (PCR) is a technique used to amplify DNA, in vitro
amplify DNA —> PCR involves making lots of copies of a specific region of DNA (usually necessary if we want to test, analyse or use a DNA sample)
in vitro —> PCR is performed in a test tube, rather than a living organism
scientists way of DNA replication
involves copying DNA
occurs in a test tube
only a specific region is copied
Materials needed for PCR
Template —> DNA sample which is going to be copied in PCR
free nucleotides —> building blocks of DNA
Heat-table DNA polymerase —> enzyme which catalyses DNA synthesis, according to the base sequence of the template
since PCR involves large temperature changes, its important that we have a DNA polymerase which can survive them - heat-stable
primers —> short single stranded pieces of chemically synthesised DNA which flank the target region
buffer —> liquid into which all of the other ingredients are added; prevents sudden pH changes
steps in PCR
put the mixture into a thermal cycler (PCR uses variations in temperature to control the replication process)
Denaturation
PCR reaction mixture is heated to 95 deg
DNA is denatured —> separates to form two single strands
Annealing
PCR reaction mixture is cooled to 55 deg
allows primers to anneal (bind) to the template DNA
extension
PCR reaction mixture is heated to 72 deg
DNA polymerase moves down the template, synthesizing new DNA
Denaturation
PCR reaction mixture is reheated to 95 deg
separates the DNA and stops DNA synthesis, so that the process can be repeated
Gel Electrophoresis
steps in Gel electrophoresis
prepare the set up
pour liquid agarose gel into a mould
insert a well comb
once the mould has set, place it into a gel box
immerse gel with buffer (can conduct electricity)

Load DNA samples and DNA ladder
each DNA sample is transferred intro its own well
a DNA ladder is added into one well
NOTE: don’t forget to record what you put in each well
Run the gel
turn on the power - a current runs though the gel
DN migrates through the gel, towards the positive pole (opposites attract)
as the gel has lots of tiny pores the small DNA fragments can fit through the pores while the larger ones can’t thus the smaller DNA fragments go through the gel faster than the larger DNA fragments


visualising the DNA fragments
add a dye to stain the DNA fragments
band —> contains a large number of DNA fragments of the same size which have ultimately travelled to the same position in the gel
applications of PCR and Gel electrophoresis
DNA sequencing —> process of determining the base sequence (ATCG) of a DNA sample
use PCR to amplify DNA
use sanger reaction to produce fragments for sequencing
run fragments on electrophoresis gel to determine sequence
DNA profiling —> process of analysing DNA variations, for the purpose of identification
use PCR to amplify short tandem repeats (STRs)
run fragments on electrophoresis gel to determine the size of the STR fragments
recombinant DNA —> DNA which contains genes from two or more different sources
PCR can be used to amplify a targets gene
run recombinant DNA on electrophoresis gel to determine whether gene has been successfully inserted
DNA Sequencing
DNA Sequencing
What’s DNA Sequencing
process of determining the sequencing of nucleotide (ATCG) in a piece of DNA
the scale of DNA sequencing varies
single gene
whole genome
steps in DNA sequencing (according to the sanger method)
gel electrophoresis
collect a DNA sample
obtained from any material which contains cells
hair, saliva, blood
extract DNA from sample
chemicals are added which break open the cells
DNA is separated from the other cell components such as proteins
Amplify DNA
involves using PCR to make lots of copies of the DNA
optional: only necessary if the sample was too small to provide enough DNA
perform sanger sequencing reaction
GOAL: separately identify the position of each nucleotide
use 4 ‘special’ PCRs = normal PCRs with a chain-terminating nucleotide

determine DNA sequence
run all 4 PCR reactions on an electrophoresis gel, to determine the lengths of the fragments in each reaction
RULES FOR DETERMINING A DNA SEQUENCE
work from smallest to largest fragment (one at a time)
identify which PCR reaction the fragment came from
the next nucleotide is complementary to the chain-terminating nucleotide in that reaction
capillary electrophoresis
collect a DNA sample
obtained from any material which contains cells
hair, saliva, blood
extract DNA from sample
chemicals are added which break open the cells
DNA is separated from the other cell components such as proteins
Amplify DNA
involves using PCR to make lots of copies of the DNA
optional: only necessary if the sample was too small to provide enough DNA
perform sanger sequencing reaction
GOAL: separately identify the position of each nucleotide
use 4 ‘special’ PCRs = normal PCRs with a florescent chain-terminating nucleotide
Determine DNA sequence
run all 4 PCR reactions on a capillary electrophoresis gel
smaller fragments move towards the end of the tube faster
as each fragment passes the laser, its fluorescent tag lights up
fluorescence is measured by a detector and recorded to produce an electropherogram
RULES FOR DETERMINING A DNA SEQUENCE
work from left to right
the next nucleotide is complementary to the fluorescent chain-terminating nucleotide
Applications of DNA sequencing
in medicine, DNA sequencing can be used to determine if a patient is at risk of a genetic disease
genetic diseases are usually associated with the presence of particular genes
an example of a genetic disease that can be identified using DNA sequencing is sickle cell anaemia
DNA sequencing is a useful tool in scientific research because it can be used to study genomes and the proteins they encode, at a molecular level
in genome mapping, DNA sequencing is used to determine he locations of genes and the distance between them
potential drug targets and variations in drug targets can be identified using DNA sequencing. this provides an opportunity for personalised medicine
DNA sequencing is used in evolutionary biology to determine inheritance patterns
evolutionary biology is the study of how different organisms are related and how they evolved
the uniqueness of a persons genetic code means that we can identify them by sequencing their DNA
this can be used to determine parentage or the identity of a culprit/victim involved in a crime.


