In Vito cloning Flashcards
1
Q
In vito gene cloning takes place ______ of a living organism.
A
Outside
2
Q
What are the requirements for PCR to occur?
A
- DNA fragment
- DNA nucleotides
- Taq polymerase (DNA polymerase)
- Primers
3
Q
What do primers do?
A
Signal where Taq polymerase should add nucleotides
4
Q
What does PCR stand for?
A
Polymerase Chain Reaction
5
Q
There are ___ stages of PCR.
A
3
6
Q
What happens in the first stage of PCR?
A
- DNA fragment is heated to a high temperature (95°C)
- DNA strands separate.
7
Q
What happens in the second stage of PCR?
A
- Temperature is reduced (55°C).
- Allows primers to join to complementary bases at the end of each strand.
8
Q
What happens in the third stage of PCR?
A
- Temperature is slightly increased (72°C).
- Taq polymerase adds complementary nucleotides to each strand.
- Two new identical DNA fragments are produced
9
Q
What is an advantage of using PCR to in vivo cloning?
A
- Takes less time (PCR takes around 2 min; in vivo cloning takes around 20 minutes for bacteria to divide)
10
Q
What is an advantage of using in vivo cloning to PCR?
A
- In vivo cloning only multiplies the DNA fragment wanted, not any contaminating DNA (restriction endonucleases are complementary to the target DNA fragments only).
- In PCR all of the DNA in the machine is multiplied, so contaminating DNA is multiplied too.
