Molecular Cloning and Gene Editing Flashcards

Flashcards covering key concepts and techniques in molecular cloning, PCR, CRISPR-Cas9, and stem cell biology.

Plasmid

Small circular DNA used as vectors in cloning; essential for gene cloning and expression.

Vector

DNA molecule that carries and replicates foreign DNA within a host cell.

Shuttle Vector

Plasmid that can replicate in two different species, facilitating gene transfer.

cDNA

Complementary DNA synthesized from RNA; lacks introns and contains only the coding region.

Competent Cells

Bacterial cells that can take up foreign DNA from their environment.

Gene Cloning

Process of inserting a gene of interest into a vector and replicating it within a host.

Site-Directed Mutagenesis

Method to introduce specific mutations at predetermined sites within a DNA sequence.

PCR (Polymerase Chain Reaction)

Technique used to amplify specific DNA sequences.

Denaturation (PCR)

The stage in PCR where double-stranded DNA is heated to separate it into single strands (94-98°C).

Annealing (PCR)

The stage in PCR where temperature is lowered, allowing primers to bind to single-stranded DNA (50-65°C).

Extension (PCR)

The stage in PCR where DNA polymerase synthesizes new DNA strands by extending primers (72°C).

Western Blotting

Technique used to detect specific proteins in a sample using gel electrophoresis and antibodies.

Restriction Endonucleases (REs)

Enzymes that cut DNA at specific sequences; essential for cloning.

Palindromic sequences

A DNA sequence where reading from the 5'-end to the 3'-end on one strand is identical to reading from the 5'-end to the 3'-end on the complementary strand

DNA Ligase

Enzyme that joins DNA fragments by forming phosphodiester bonds.

Blue-white screening

Method to identify cells that have successfully taken up recombinant DNA; disruption of the LacZ gene results in white colonies.

CRISPR-Cas9

Gene-editing tool that uses Cas9 to cut DNA at a specific location guided by RNA.

Donor DNA (CRISPR)

The template used to introduce specific changes during the CRISPR process.

Guide (spacer) RNA (CRISPR)

Guides the Cas9 enzyme to the target DNA sequence for editing.

Totipotent Cells

Can differentiate into any cell type, including extra-embryonic tissues.

Pluripotent Cells

Can give rise to almost all cell types but not extra-embryonic tissues.

Multipotent Cells

Limited to differentiating into a few cell types.

Unipotent Cells

Can only produce one cell type.