Flashcards: Molecular Techniques in DNA Amplification — 20 cartões

DNA amplification — in vitro method?

Cell-free, rapid DNA copying technique.

PCR — main process steps?

Denaturation, annealing, extension.

Primer design — importance?

Ensures specificity and efficiency.

qPCR — detection method?

Fluorescent probes monitor amplification.

DNA cloning — process?

Transfer DNA into cells for proliferation.

Non-selective amplification — purpose?

Amplifies all DNA fragments without targeting.

PCR data analysis — method?

Real-time fluorescence during exponential phase.

LAMP — temperature?

Constant, around 60-65°C.

DNA polymerase — role?

Synthesizes new DNA strands.

PCR — advantage?

Fast, specific, high throughput.

PCR — limitation?

Error rate, size limits, contamination risk.

Taq polymerase — source?

From Thermus aquaticus bacteria.

Taq — proofreading?

Lacks 3' to 5' exonuclease activity.

Fidelity enzyme — alternative?

Archaeal polymerases with proofreading.

PCR — main advantage?

Rapid, sensitive DNA amplification.

PCR — main limitation?

Error rate and contamination risk.

DNA cloning — large fragment?

Up to 1MB in bacteria.

Non-selective amplification — key tool?

Adaptor oligonucleotides with universal sequences.

qPCR — quantification?

Absolute via standard curve; relative via ΔΔCt.

LAMP — primer count?

4 to 6 primers recognizing multiple regions.