Principles of Molecular and Immunologic Studies - Amplification Techniques

These flashcards cover key terminology and concepts related to amplification techniques in molecular and immunologic studies.

Reverse Transcription

The process through which a single-stranded RNA molecule gives rise to a complementary DNA (cDNA) molecule.

RT-PCR

Reverse Transcription Polymerase Chain Reaction, a method used to amplify and analyze the cDNA derived from RNA.

cDNA

Complementary DNA synthesized from a single-stranded RNA template through reverse transcription.

Primer-dependent polymerase reaction

A molecular technique that requires primers for the synthesis of DNA from an RNA template.

Two-step RT-PCR

A method involving two main steps: producing and purifying cDNA followed by amplification using PCR.

Real-time PCR

A quantitative PCR method that monitors the progress of PCR as it occurs to measure DNA.

Threshold cycle (CT)

The cycle number at which the fluorescence of the PCR product surpasses the threshold level.

SYBR Green

A DNA-specific dye that binds to double-stranded DNA and emits fluorescence when excited.

Taq polymerase

A thermostable DNA polymerase enzyme used in PCR to amplify DNA.

Fluorescence Resonance Energy Transfer (FRET)

A technique that measures the transfer of energy from a donor fluorophore to an acceptor fluorophore.

TaqMan assay

A PCR technique using a specific probe that emits fluorescence upon probe cleavage by Taq polymerase.

Molecular Beacon

A probe that fluoresces when bound to the target DNA during PCR amplification.

Melting Curve Analysis

A technique used to analyze the melting behavior of DNA, providing information about the DNA's properties.

Exponential growth phase

The phase in PCR where the amount of product doubles with each cycle.

Plateau phase

The phase in PCR where the reaction reaches a maximum product concentration and amplification slows.

Quantitative PCR (qPCR)

A method for measuring the quantity of target DNA by monitoring the amplification process in real-time.

Master mix

A pre-prepared solution containing all the necessary components for a PCR reaction, excluding the template.