Polymerase Chain Reaction (PCR)
A technique that allows researchers to produce a large number of identical DNA molecules in vitro
PCR first step
Denaturation 95' C - Break hydrogen bonds
PCR second step
Priming 60' C - adding RNA primase (3' hydroxyl)
PCR third step
Extension 72' C - copying
Machine that automates the repetitive process of PCR
Technique used to separate molecules by size shape and electrical charge.
Technique that allows researchers to stabilize specific DNA sequences from an electrophoresis gel and then localize them using DNA dyes or probes.
Technique that is used to detect RNA
Numerous distinct ssDNA molecules bound to a substrate and used to probe for complementary sequences.
Natural DNA inserting methods
Transformation; Transduction; Conjucation
Artificial DNA inserting methods
Electroporation; Protoplast fusion; Injection (gene gun and microinjection)
Use of an electrical current to puncture microscopic holes through a cell's membrane so that DNA can enter the cell from the environment.
When protoplasts encounter one another, their cytoplasmic membranes may fuse to form a single cell that contains the genomes of both "parent" cells creating a recombinant cell.
Injection: Gene gun
blank .22 caliber cartridge or compressed gas fire tiny tungsten or gold beads coated with DNA into a target cell.
A geneticist inserts DNA into a target cell with a glass micropipette having a tip diameter smaller than that of the cell or nucleus.
Fluorescence in situ hybridization (FISH)
When researchers know of a particular gene exclusive to that organism, they can locate the gene by attaching fluorescent chemicals to short, single strands of nucleic acid molecules that are complementary to the gene or it's transcribed mRNA.
Determining the exact order of the base pairs in a segment of DNA
Pharmaceutical and Therapeutic Applications
Protein synthesis; vaccines; genetic screening; DNA fingerprinting; Gene therapy; Medical diagnosis; Xenotransplants
Production of transgenic organisms; Herbicide tolerance; Salt tolerance; Freeze resistance; Pest resistance; Improvements in nutritional value and yield;