Gene Tech - In Vitro Flashcards
1
Q
what is in vitro cloning?
A
(Outside the body)
Using PCR
2
Q
What is the purpose of PCR?
A
To copy DNA fragments
3
Q
What does PCR require?
A
- DNA fragment to be copied
- TAQ polymerase: Resistant to high temperatures, used to join together nucleotides
- Primers
- Nucleotides
4
Q
What are the stages of PCR?
A
- Separation of the DNA strand
- Addition of primers
- Synthesis of DNA
5
Q
What happens during the separation stage of PCR?
A
- Temperature increases to 95°C causing the double helix DNA strand to separate due to the breaking of hydrogen bonds.
6
Q
What happens in the addition stage of PCR?
A
- Mixture is cooled to 55°C, causing the primers to anneal (join) to their complementary bases.
- Primers provide the starting sequence for DNA polymerase.
- Also prevent the two separate strands from re-joining.
7
Q
What happens during the synthesis stage of PCR?
A
- Temp increases to 72°C.
- Optimum temp for DNA polymerase to add complementary nucleotides along each separated DNA strand until it reaches the end strand.
8
Q
What happens after PCR?
A
Process repeats
9
Q
Advantages of in vitro gene cloning
A
- Many copies of DNA can be made in a short amount of time but also contaminate DNA
- Only samples are required, no living organism
