Topic8 In Vitro Cloning Flashcards
What does In Vitro mean?
In Vitro means outside living organisms
Once DNA fragments been isolated what needs to happen?
Once DNA fragments been isolated it needs to be cloned to create large quantities so
Amplifying DNA Fragments- and this is done in Vivo and in Vitro
Where does Amplifying DNA fragments happen In Vitro?
The Polymerase Chain Reaction (PCR)
Fragments of DNA in Vitro is amplified by PCR
What is PCR ?
Polymerase Chain Reaction
What is the special enzyme used in PCR?
What is the properties?
And where is it found?
Enzyme DNA TAQ polymerase.
Found in hot temp and optimum temperature is 72 Celsius
Denature over 100 Celsius
What is the first stage in PCR method?
The temperature first increased to 95 Celsius breaks hydrogen bonds and splits DNA into single strands
DENATURING
What is the Second Stage in PCR Method?
Temperature then decreased to 55 Celsius so that Primers can attach
Annealing
What is the thirds stage in PCR method?
Enzyme DNA polymerase joins complementary free nucleotides and make new stand to align next to each template
Synthesis
What is the last stage in PCR method?
Temperature increased to 72 Celsius for this stage
The optimum for Enzyme DNA TAQ polymerase
What are all the stages in PCR method?
1.Denaturing
2.Annealing
3.Synthesis
What are the advantages of PCR?
Automated- more efficient
Rapid
Doesn’t require living cells- quicker and less complex technologies needed
