Nucleic Acid Amplification and PCR

Flashcards covering key concepts and vocabulary related to Nucleic Acid Amplification and the Polymerase Chain Reaction (PCR).

PCR

Polymerase Chain Reaction; a technique used to amplify DNA.

Amplicon

Each individual copy produced by the PCR process.

Master Mix

A solution containing dNTPS, DNA polymerase, primers, and buffer used in PCR.

Buffers

Solutions that maintain a consistent pH, specifically Tris (pH 8.4), MgCl2, and KCl in PCR.

Primers

Short DNA sequences that bind to the template DNA and are necessary for DNA synthesis during PCR.

Denaturation

The first step in PCR where the temperature is raised to break hydrogen bonds in DNA.

- lasts 20 to 60 seconds

- raised to 90 to 96 degrees

Annealing

The step in PCR where the temperature is lowered to allow primers to bind to the template DNA.

- raised 50 to 70 degrees

- lasts 20 to 90 seconds

Extension

DNA polymerase adds dNTPS and elongs the chain

- raised 68 to 75 degrees

- lasts 10 to 60 seconds

Cycle

One complete round of denaturation, annealing, and extension in PCR.

Gel Electrophoresis

A method used to assess the end product of PCR by separating DNA fragments based on size.

Misprime

An error during the annealing step when a primer binds to an incorrect sequence.

Primer Dimers

Formed when two primers bind to each other instead of their intended target on the DNA.

Continuous amplification

Continuous amplification of template by repeating cycles of replication

-  Total product = 2^N (2 raised to the number of cycles youve completed)

Total product calculation

The total amount of DNA generated after N cycles of PCR is calculated as 2^N.