chapter 6 Flashcards
1
Q
Polymerase Chain Reaction
A
Amplify (increase) amount of DNA from a few to millions
- denature
- anneal (primers)
- extension
2
Q
Ingredients
A
- Reaction buffer
- dNTPS
- MgCl2
- Primers
- Polymerase
3
Q
coding strand
A
- same as RNA transcript
- sense strand
- non template
4
Q
non coding strand
A
- template strand
- anti sense
5
Q
forward primer (5-3)
A
- coding
- anneals to template and same as og coding
6
Q
reverse primer (3-5)
A
- non coding
- anneals coding = identical to og non coding
7
Q
conditions
A
The 5ʹ end can tolerate mismatched bases, but the 3ʹ end cannot.—must be anneal properly
20-30 long
50-60 melting for primer
8
Q
needs to be between
A
1 min and 2-5 degrees
9
Q
PCR cycling conditions
A
need to know length of amplicon and primer temp
