In Vitro Gene Cloning

Question 0

What components are needed?

Answer 0

DNA Polymerase 
Primers
Thermocylcer 
DNA fragments
Nucleotides

Question 1

Describe PCR

Answer 1

Polymerase chain reaction

Automated way of copying DNA fragments using a thermocylcer

Question 2

What temp is the thermocylcer heated to, to seperate the DNA fragment into 2?

Answer 2

95°c

Question 3

What is the temp cooled to in the second stage?

And what does this allow for?

Answer 3

Cooled to 55°c

Allowing for primers to anneal to complementary bases at the end of the DNA fragment

Question 4

What do the attached primers provide?

Answer 4

The starting sequence for DNA polymerase to begin DNA copying as polymerase can only attach nucleotides to the end of an existing chain
Also prevent the two separate DNA fragments strands from rejoining

Question 5

Why is the temp increased to 72°c?

Answer 5

This is the optimum temp for DNA polymerase to add complementary nucleotides along each of the separate DNA stands

Question 6

Where do nucleotides start and finish being added?

Answer 6

Nucleotides are attached at the primers on both strands and are added in sequence until the end of the chain has been reached

Question 7

2 advantages of In Vitro gene cloning?

Answer 7

Extremely rapid - billions of copies can be made in hours, particularly valuable when only a small amount of DNA is available
Does not require living cells - all that’s required is a base sequence of DNA that needs amplification