3 5 Genetic Modification Gelelectrophoresis 10

Lab technique used to separate and isolate

and Biotechnology protein or DNA
fragments into mass size
sampler are placed in block ofget and an electric
Polymerase chain reaction V 1 current is
applied sampler move through gel
PCR artificial method for DNA replication small sampler move fatter samplerseparate by size
mass
stages 1
Agarre Gel Electrophoresis DNA
1 Denaturation DNA sample is heated to DNA is cut into fragments by restriction endonuclease
be separated into 2 strands enzyme separating them into diff lengths
2 DNA primers attach DNA sampler are placed in agarre get and fragment
Annealing
3 Elongation
Taq Cheat tolerant DNA size is calculated
polymerase binds to primer and copier strand southern blotting specific sequencer are identified by
radio labelling then transferring them to a membrane to vinalize via
auto radiography in 3 2

2
Polyacrylamide Gel Electrophoresis Protein
Proteins are treated with anionic detergent SDS linearise and make a
uniform negative charge
protein sampler are placed in polyacrylamide gel and sizes are compared
western Blotting separated proteins are transferred into membrane and target
protein are identified with specific monoclonal antibodies
DNA profiling IV 3 Gene Transfertgere modification V4
Technique by which individuals are identified and Gere determines specific trait by encoding in a
compared via their DNA profiler protein
In non coding DNA regions DNA not activated by since genetic code is Almost universal an organism
body satellite DNA made of STR's con express a new trait if appropriate gene is induced
people have dif STR repetition this creates Newly genetic modified organism transgenic
unique DNA profile
vector D NA
Steps A 1 molecule wed to
DNA sample blood saliva semen is taken and carry intrez gene
amplified by PCR into foreign cell
satellite DNA is Cut with restriction enzymes o Ex plasmids vives
generate fragments separated by gel electro
profiles are compared Process A 2
Forensics murder weapon matches killer 1
Isolating gene and vector
Paternity test fragments from child are Gene of interest is amplified via PCR
either from mom or dad Vector wed Bacterial plasmids 13.2 autonomos
5 2 el f replication and
expresio es

aman

amo
 2 Digestion with restriction enzymes Selection and expression
TO incorporate rector into gere both matte cat with Recomb construct is introduced into into organism
restriction enzymes at recognition sites plasmid vector contain antibiotic resistance gene only
Restriction enzymes cut the DNA to make blunt transgenic cells will grow new gene will start expressing
or sticky ends better
GMO Debate A 3
amo genetically modified organism
AMO s are beneficial and
harmful to
Human health
Agriculture
Environment

3 Ligationof rector and INEA

Gene is inserted into plasmid rector
that her been cut with same retriction
endonuclease enzyme clones U s
sticky end of gene and vector overlap clones genetically identical organisms group of cells derived from single
by complementary bare pairing clones asexual reproduction
parent cell
Gene and vector ave ligared by sexual reproduction identical twin
DNA ligare enzyme to clone multicellular organisms
stem cells somatic cell Nuclear Transfer SINT
requires

SCNT method which cloned embryos con be produced

by wing
differentiated adult cells

recombinant construct
 Cloned embryos via MNT A 4 3 Fragmentation starfish annelid worm
somatic cells are removed from adult and cultured New organism grow from separated fragment of parent
Un fertilised
egg is removed from adult female its organism
haploid nuclear is removed e nucleated
egg cell
E c c is fired with adult donor nuclear diploid with 4 Parthenogenesis Irme insects fin reptiles amphibians
donor DNA Embryos formed from un fertilised egg
An electric current stimulates the egg to develop
into an embryo implanted into surrogate and
will grow into adult donor genetic clone

plant cloning method

plants have capacity of vegetative propagation small
pieces con be induced to grow independently
Natural Cloning v 6
Species with asexual reproduction natural All roots and shoots are capable of L
bacteria fungi some plants animals cloning garlic onion bulbs underground stems runner
some plants reproduce asexually via pores
Animal cloning methods algae fem
1 Binary fission bacteria protists
parent organism divider equally in 2 2 genetically
identical daughter
2 Budding
year
cells split from parent organism smaller
daughter organism which split from parent
 Human cloning method
Identical twin monozygotic Zygote splits into 2
identical cells
Non identical twin dizygotic un fertilised
egg
splits into 2 cells

stem cutting 5 1
separated portion of plant cell that
regrow into an independent clone
This is a common method to
rapidly
propagate plant species