Amplifying The Blueprints Of Biology Flashcards
0
Q
PCR steps
A
- denaturation (92-95)
- primer annealing (50-60)
- extension (70-72)
1
Q
PCR reagents
A
Template DNA Primers dNTPs MgCl Standard buffer Taq Water
2
Q
Tm
A
4(G+C)+2(A+T)
3
Q
Applications of PCR
A
DNA fingerprinting Cycle sequence MPLA Quantitative PCR Micro array analysis Cloning Mutation detection Genotyping
4
Q
Old-school SDM
A
Primer contains desired mutation
After denaturation, primer annuals to target DNA
Product is introduced into host cell and cloned
Get a mixture of wild-type and mutant
5
Q
Quick-change SDM
A
- both primers contain mutation
- complementary mutagenic strands generated
- parental DNA is digested with Dpn1 to target methylated DNA
6
Q
Tasman assay
A
5’fluorophore and 3’quencher
When primer extends, the fluorophorw is released from quencher
