Amplifying The Blueprints Of Biology Flashcards Preview

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Flashcards in Amplifying The Blueprints Of Biology Deck (7):
0

PCR reagents

Template DNA
Primers
dNTPs
MgCl
Standard buffer
Taq
Water

1

PCR steps

- denaturation (92-95)
- primer annealing (50-60)
- extension (70-72)

2

Tm

4(G+C)+2(A+T)

3

Applications of PCR

DNA fingerprinting
Cycle sequence
MPLA
Quantitative PCR
Micro array analysis
Cloning
Mutation detection
Genotyping

4

Old-school SDM

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

Quick-change SDM

- both primers contain mutation
- complementary mutagenic strands generated
- parental DNA is digested with Dpn1 to target methylated DNA

6

Tasman assay

5'fluorophore and 3'quencher
When primer extends, the fluorophorw is released from quencher