Polymerase Chain Reaction (PCR) Flashcards
1
Q
Define
Polymerase Chain Reaction (PCR)
A
method to amplify DNA in vitro using many cycles of DNA denaturation, primer annealing, and DNA polymerization with a heat-stable polymerase
2
Q
Step 1 of PCR
A
94°C for 5 minutes; denaturation d/t heat separates DNA strands, exposing the nucleotide bases
3
Q
Step 2 of PCR
A
48°C for 60 seconds; DNA primers hybridize to the exposed strands
4
Q
Step 3 of PCR
A
72°C for 60 seconds; DNA polymerase begins replication of the targeted regions (starting at the primers)
5
Q
What is the general purpose of PCR?
A
amplify a target gene of a given DNA strand for the purposes of ID
6
Q
How many times are steps 2-4 of PCR repeated?
A
steps 2-4 repeated 30 times
7
Q
What is the target of amplification for class?
A
16S ribosomal subunit
8
Q
PCR Components
(5 total)
A
- buffer solution (MgCl2 and pH buffers)
- DNA template to be copied
- many copies of forward and reverse primers
- free DNA nucleotides
- DNA polymerase enzymes
