Lecture on Polymerase Chain Reaction

These flashcards cover key concepts related to Polymerase Chain Reaction (PCR), including definitions and important parameters in primer design.

Polymerase Chain Reaction (PCR)

A technique developed in the 1980s for amplifying a specific sequence of DNA.

Denaturation

The first stage of the PCR cycle where the DNA is melted at approximately 95°C.

Annealing

The second stage of the PCR cycle where primers bind to the template DNA at 60-65°C.

Extension

The third stage of the PCR cycle where DNA polymerase synthesizes new DNA strands at 68-72°C.

Taq DNA Polymerase

A type of DNA polymerase isolated from Thermus aquaticus, used in PCR for its heat stability.

Thermocycler

An instrument used to perform PCR by cycling through various temperature stages.

ΔG (Delta G)

Represents the stability of primers; more negative values indicate higher stability.

Hairpin Loop

A secondary structure formed when a primer folds back on itself, which is undesirable.

Self-Dimers

Formed when a single primer binds to itself; not desired for effective PCR.

GC Clamp

A short stretch of G/C at the 5’ end of a primer which increases stability and specificity.

Cross-Dimers

Dimers formed between forward and reverse primers; stability should be minimized.

Sticky Primer

A primer with strong binding at the 5' end, leading to increased stability.

Non-Sticky Primer

A primer with less binding at the 3' end to allow proper extension by DNA polymerase.