e1-2 Flashcards
Blue Word
Definition
PCR
Polymerase Chain Reaction; a method to amplify specific DNA sequences in vitro.
primers
Short sequences of nucleotides that provide a starting point for DNA synthesis.
Taq Polymerase
A heat-stable DNA polymerase from Thermus aquaticus used in PCR.
Denaturation
The step in PCR where double-stranded DNA is heated to separate strands.
Annealing
The step in PCR where primers bind to the template DNA.
Extension
The step in PCR where Taq polymerase adds nucleotides to synthesize new DNA.
Gel Electrophoresis
A technique to separate nucleic acids or proteins based on size using an electric field.
Ladder
A molecular weight marker used in gel electrophoresis to estimate DNA fragment sizes.
Fluorescence microscope
A microscope that uses fluorescence to visualize specific molecules.
fluorophore
A molecule that absorbs and emits light at specific wavelengths.
in vivo
Experiments conducted within a living organism.
in vitro
Experiments conducted outside a living organism, usually in a test tube or culture dish.
kDa
Kilodalton, a unit of molecular weight for proteins.
transfer
The process of moving nucleic acids or proteins from a gel onto a membrane for further analysis.
Southern blot
A technique to detect specific DNA sequences on a membrane.
Northern blot
A technique to detect specific RNA sequences.
Western blot
A technique to detect specific proteins using antibodies.
antibody
A protein that binds to specific antigens for immune response or research purposes.
antigen
A molecule recognized by an antibody, usually foreign.
hybridization
The binding of complementary nucleic acid strands.
SDS-PAGE
A technique that separates proteins based on size using a gel and electric field.
reverse transcription
The process of making DNA from RNA using reverse transcriptase.
RT-PCR
Reverse transcription PCR, used to amplify RNA by converting it into DNA first.
in situ hybridization
A technique to visualize DNA or RNA within cells or tissues.
GFP transcriptional fusions
A technique where GFP is placed under a gene’s promoter to study gene expression.
GFP translational fusions
A technique where GFP is fused to a protein to track its location.
immunohistochemistry
A technique using antibodies to detect proteins in tissue sections.
immunoprecipitation
A technique to isolate a specific protein from a mixture using antibodies.
trans-gene
A gene artificially transferred into an organism.
reporter gene
A gene used to measure gene expression, like GFP.
restriction enzyme
An enzyme that cuts DNA at specific sequences.
plasmid
A small circular DNA molecule used in genetic engineering.
transformation
The process of introducing foreign DNA into a cell.
recognition sites
Specific DNA sequences where restriction enzymes cut.
restriction mapping
A technique to determine the locations of restriction enzyme cut sites.
Multiple Cloning Site (MCS)
A region in a plasmid with several restriction sites for cloning.
Origin of Replication (ori)
The sequence where DNA replication begins in a plasmid.
selectable marker
A gene that allows selection of successfully transformed cells.
blunt ends
DNA fragments with no overhangs after restriction enzyme digestion.
5’ overhanging sticky ends
Single-stranded DNA overhangs at the 5’ end after restriction digestion.
3’ overhanging sticky ends
Single-stranded DNA overhangs at the 3’ end after restriction digestion.
selection
Process of identifying cells that have taken up foreign DNA.
