Polymerase Chain Reaction and Molecular Phylogeny

These flashcards cover key concepts regarding the Polymerase Chain Reaction (PCR), the use of 16S rRNA in microbial comparison, and the basics of molecular phylogeny.

Polymerase Chain Reaction (PCR)

A method used to amplify DNA sequences.

16S rRNA

A gene used as a marker for comparing microbial species.

Marker genes

Genes used to identify and classify organisms based on genetic similarities.

Archaea

One of the three domains of life, characterized by prokaryotic microorganisms.

Bacteria

One of the three domains of life, consisting of prokaryotic microorganisms.

Eukarya

One of the three domains of life, consisting of organisms with eukaryotic cells.

Molecular phylogeny

The study of evolutionary relationships between species based on molecular data.

Molecular Clock

A method used to estimate the time of evolutionary events based on genetic mutations.

Taq Polymerase

An enzyme used in PCR that synthesizes new DNA strands.

dNTP

Deoxynucleoside triphosphates, the building blocks for new DNA synthesis.

Tris-HCl

A buffer used to maintain pH in PCR reactions.

EDTA

A chelating agent used in PCR to keep DNA intact.

MgCl2

A cofactor required for the activity of Taq polymerase.

Thermal Cycler

An instrument used to temperature-manage PCR amplification.

PCR Steps

The three steps of PCR are denaturation, annealing, and extension.

Initial Denaturation

The first step of PCR where DNA is heated to 95°C for 3 minutes to separate strands.

Annealing

The step in PCR where primers bind to the target DNA at a lower temperature.

Extension

The step in PCR where Taq polymerase synthesizes new DNA strands.

Final Extension

The concluding step of PCR where the reaction is held at 72°C for a short period to finalize DNA synthesis.