Topic8 In Vitro Cloning

Question 1

What does In Vitro mean?

Answer 1

In Vitro means outside living organisms

Question 2

Once DNA fragments been isolated what needs to happen?

Answer 2

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

Question 3

Where does Amplifying DNA fragments happen In Vitro?

Answer 3

The Polymerase Chain Reaction (PCR)

Fragments of DNA in Vitro is amplified by PCR

Question 4

What is PCR ?

Answer 4

Polymerase Chain Reaction

Question 5

What is the special enzyme used in PCR?
What is the properties?

And where is it found?

Answer 5

Enzyme DNA TAQ polymerase.

Found in hot temp and optimum temperature is 72 Celsius

Denature over 100 Celsius

Question 6

What is the first stage in PCR method?

Answer 6

The temperature first increased to 95 Celsius breaks hydrogen bonds and splits DNA into single strands

DENATURING

Question 7

What is the Second Stage in PCR Method?

Answer 7

Temperature then decreased to 55 Celsius so that Primers can attach

Annealing

Question 8

What is the thirds stage in PCR method?

Answer 8

Enzyme DNA polymerase joins complementary free nucleotides and make new stand to align next to each template

Synthesis

Question 9

What is the last stage in PCR method?

Answer 9

Temperature increased to 72 Celsius for this stage

The optimum for Enzyme DNA TAQ polymerase

Question 10

What are all the stages in PCR method?

Answer 10

1.Denaturing

2.Annealing

3.Synthesis

Question 11

What are the advantages of PCR?

Answer 11

Automated- more efficient

Rapid

Doesn’t require living cells- quicker and less complex technologies needed