21) Manipulating Genomes Flashcards
What do you add in a solution when finding a DNA Profile?
- Free nucleotide bases
- Primers
- DNA Sample
- DNA Polumerase
Define PCR
Why is this important
Polymerase Chain Reaction
Amplify and produce millions of copies of a fraagment of DNA
Describe Denaturing stage in PCR
Mixture heated to 95 degress to break H bonds
Why is DNA Polymerase efiicient even at high temperatures
They don’t denature , many cycles of PCR can be carried out without using new enzymes
Describe what happens in annealing stage of pcr
Cooled between 50 - 60 degrees .
Where the primers bind to the strands
Describe what happens in the extension stage of pcr
Heat up to 72 degrees.
Dna polymerase work and lines up free nucleotides along template strang. COmplementary base pairing forms complementary strands.
this creats 2 new copies
How does electrophoresis contribute in a DNA Profile
Add a small current through the gel.
DNA is negatively charged , they are placed at the -ve electrode
move toeards the positive electrode
smaller fragments travel faster than longer.
What is the role of restriction enzymes
Cut fragments at restriction sites,seperate specific fragments to the rest of the DNA.Create sticky ends, used to anneal complementary sequences,
Tell me how to create a DNA Profile
DNA is isolated , add restriction enzyme to get fragment needed.
Pcr to amplify the fragments.
Run onto electrophoresis gel and allow small current to pass.
Include a type of fluorescent dye and place under UV light .
Give examples on where we us DNA Profiling
Medical diagnosis , forensic science , ability to diagnose genetic disorders.
Descrribe how you can genetically engineer for a gene you want
1) Isolate required gene by using restiction enzymes.
2) Isolated DNA is inserted into a vector using DNA ligase.
3) Sticky ends from the reuired DNA and Vector DNA join by complementary base pairing by DNA ligasse .
4) Reform sugar-phosphate backbone by ligation
5) Now create recombinant DNA
6) Recombinant DNA transfer into bacterial cell by electroporation
7) THhis is called transformation
What is the significance of electroporation
Increase the permeabbility of the bacterial cell membranes to allow to take in plasmids
+ev in GE be used to create insect-resistance in plants
Reduce amount of chemical pesticides whcih can harm the environment
-ve in GE used to create insec-resistance in plants
Encourage monoculture , decrease in genetic variation , more susceptible/ genetically identical to the same diseases.
+ve in GE in producing Drugs from animals
be made in large quantities , available to more people
