Recombinant DNA technology
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Created by:
Portsandie on April 28, 2012
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19 terms
Terms | Definitions |
|---|---|
 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 |
| Thermocycler | Machine that automates the repetitive process of PCR |
| Gel electrophoresis | Technique used to separate molecules by size shape and electrical charge. |
| Southern Blot | Technique that allows researchers to stabilize specific DNA sequences from an electrophoresis gel and then localize them using DNA dyes or probes. |
| Northern Blot | Technique that is used to detect RNA |
| DNA Microarrays | 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) |
| Electroporation | Use of an electrical current to puncture microscopic holes through a cell's membrane so that DNA can enter the cell from the environment. |
| Protoplast fusion | 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. |
| Injection: Microinjection | 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. |
| DNA sequencing | 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 |
| Agricultural Applications | Production of transgenic organisms; Herbicide tolerance; Salt tolerance; Freeze resistance; Pest resistance; Improvements in nutritional value and yield; |
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