BMSC 240 Laboratory Techniques Lecture 21: Module 9: Cutting, Cloning and Isolating DNA

These flashcards cover key vocabulary terms and concepts related to the techniques discussed in the BMSC 240 Laboratory Techniques lecture on Cutting, Cloning, and Isolating DNA.

Plasmid Extraction

The process of isolating plasmid DNA from bacterial cells, typically performed using a miniprep technique.

Plasmid DNA Precipitation

The procedure of adding salt and alcohol to a solution to precipitate DNA, facilitating its collection.

Multiple Cloning Site (MCS)

A region in a vector containing several restriction sites, allowing the insertion of foreign DNA fragments.

Blue-White Screening

A method to identify recombinant bacteria based on the color change of colonies when using X-gal.

Taq Polymerase

A heat-stable DNA polymerase used in PCR that synthesizes new strands of DNA at high temperatures.

Denaturation

The first step in PCR where the double-stranded DNA is heated to separate into single strands.

Annealing

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

Extension

The step in PCR where Taq polymerase synthesizes a new DNA strand from the template.

Miniprep

A small-scale plasmid extraction from bacterial cultures, typically involving a few milliliters of culture.

dNTPs (Deoxynucleotide Triphosphates)

The building blocks of DNA used in PCR, consisting of adenine (A), thymine (T), cytosine (C), and guanine (G).

PCR Cycle

A series of temperature changes (denaturation, annealing, extension) that amplify a specific DNA region.

Error Rate of Taq Polymerase

The frequency of incorrect base incorporation, approximately one wrong base every 100,000 nucleotides.

Heat Shock

A technique used in transformation that involves exposing cells to a high temperature to facilitate the uptake of plasmid DNA.

PCR Master Mix

A prepared mixture containing all necessary reagents for PCR except for the template DNA.

DNA Lysis

The process of breaking down cell membranes and DNA to release plasmid or genomic DNA for extraction.

Phenol/Chloroform Extraction

A method used to purify DNA by separating it from proteins and other cellular debris.