Ch 2.2 PCR Flashcards
1
Q
What’s the use of PCR?
A
To copy dna in vitro( test tube)
2
Q
What does amplifying DNA in PCR mean?
A
Repeatedly copying section dna
3
Q
What is so good about PCR?
A
Easy to use
4
Q
The key components of PCR?
(4)
Minor components (2)
A
Key
- DNA template
- primers
- dNTPs ( nucleotides)
- Dna Polymerase
Minor
- water
- buffer
5
Q
What does template dna do in PCR (2)
A
- usually double stranded dna that has the section of interest
- usually larger then desired section
6
Q
What do the dna primers do in PCR?
A
- also called olignucleotide primers cuz oliglo refers 15-30 bases
- each primer anti parallel/ complementary
- located just outside desired part
7
Q
What does dNTP stand for
A
Deoxynucleoside triphosphate
8
Q
What direction are new nucleotides added in PCR?
A
5’-3’ on the developing strand
The template strand is read 3’-5’
9
Q
Will the amplified dna in PCR include the primer?
A
1) yes
10
Q
DNA pol in PCR?
A
- need to be heat resistant so use Tac polymerase ( hot spring bac)
- reads dna 3-5 add 5-3 ( dna elongation )
- need free free OH from primer to add nucleotides
11
Q
PCR amplification cycles
(3)
A
- Denaturation ( temp 95)
- Annealing ( temp 55-60)
- Extension (temp 72)
