Laboratory Techniques in DNA Manipulation

A series of vocabulary flashcards for reviewing DNA manipulation and cloning techniques.

DNA Cloning

A technique used to create copies of a specific DNA sequence.

Ligation

The process of joining two DNA strands together by forming phosphodiester bonds.

Transformation

Inserting foreign DNA into a host cell, often bacteria, to induce expression of the gene.

pBluescript

A commonly used plasmid vector for cloning and expression of genes.

GST Gene

Glutathione S-transferase gene, often used for protein expression and purification.

Colony PCR

A method to identify bacteria that contain an inserted plasmid by amplifying DNA from colonies.

SDS-PAGE

A technique for separating proteins based on their size using gel electrophoresis.

Restriction Digest

The process of cutting DNA into fragments using specific enzymes called restriction enzymes.

Miniprep

A quick method for extracting and purifying plasmid DNA from bacteria.

PCR

Polymerase Chain Reaction, a technique used to amplify a specific DNA sequence.

Spin Column

A method used for purifying DNA or RNA from a solution.

E. coli

A common bacterium used as a host for cloning and expressing recombinant DNA.

Ligase

An enzyme that facilitates the joining of DNA strands together.

Ori

Origin of replication, a sequence of DNA at which replication begins.

Ampicillin Resistance Gene

A gene providing resistance to the antibiotic ampicillin, typically used as a selectable marker.

Promoter

A DNA sequence that initiates the transcription of a gene.

X-Gal

A substrate used for detecting beta-galactosidase activity, indicating successful cloning.

IPTG

Isopropyl β-D-1-thiogalactopyranoside, an inducer of gene expression.

Heat Shock

A method used to introduce plasmid DNA into bacterial cells by temporarily increasing temperature.

Transformation Efficiency

The measure of how many cells take up foreign DNA during transformation.

Blue/White Screening

A method to identify recombinant colonies based on the color of the colonies grown on X-gal plates.

Gene Expression

The process by which information from a gene is used to synthesize a functional gene product, often proteins.

Ligated DNA

DNA that has been joined together through a ligation reaction.

Colony Selection

Choosing colonies of bacteria that contain the desired plasmid for further analysis.

Bacterial Transformation

The process by which bacteria take up foreign DNA from their environment.

Purification Process

Steps taken to isolate specific DNA or proteins from a mixture.

Multiple Cloning Site (MCS)

A region in a plasmid that contains multiple restriction sites for the insertion of DNA.