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Manufacturing pro
| Term | Definition |
|---|---|
| Amino Acid | The subunit of protein |
| Ampicillin | A penicillin that is effective against gram-negative and gram- positive bacteria. |
| Chromatography | A process in which a chemical mixture carried by a liquid or gas is separated into components as a result of differential distribution of the solutes as they flow around or over a stationary liquid or solid phase. |
| Column Chromatography | The substance to be separated are introduced onto the top of a column packed with an adsorbent, pass through the column at different rates that depend on the affinity of each substance for the solvent mixture, and are usually collected in solution as they |
| Gel Electrophoresis | The separation of nucleic acids or proteins, on the basis of their size and electrical charge, by measuring their rate of movement through an electric field in a gel. |
| Genetic Engineering | The direct manipulation of genes for practical purposes. |
| Hydrophilic | (water loving), materials attract, absorb, or dissolve in water due to polar properties. |
| Hydrophobic | (water fearing) materials repel water, forming droplets on surfaces due to nonpolar properties. |
| Insulin | A protein hormone synthesized in the pancreas that regulates blood sugar levels by facilitating the uptake of glucose into tissues. |
| Plasmid | A small ring of DNA that carries accessory genes separate from those of the bacterial chromosome. |
| Polyacrylamide | Used as a gel material in vertical electrophoresis; used to separate smaller molecules, like proteins or very small pieces of DNA or RNA. |
| Recombinant DNA | A DNA molecule made in vitro with segments from different sources. |
| Transformation | The uptake and expression of foreign DNA by a cell. |
| Transformation Efficiency | A measure of how well cells are transformed to a new phenotype. |
| Aqueous environment | Proteins fold so that the hydrophobic amino acids "hide" on the inside. |
| SDS (sodium dodecyl sulfate) | Soap that dissolves hydrophobic molecules (like GFP) |
| PAGE (polyacrylamide gel electrophoresis) | The method used to check for protein purity. |
| DNA electrophoresis | Uses agarose gel, requires PCR of DNA first, gel is loaded horizontally, DNA moves across the gel. |
| Protein electrophoresis | uses polyacrylamide gel, requires SDS application to protein 1st, gel is loaded vertically, protein moves down the gel. |
| GFP gene | Codes for GFP protein which glows green when exposed to UV light. |
| araC | gene regulates the transcription of GFP |
| Ori | allows plasmid replication |
| Bla | An ampicillin resistant gene |
| Electroporation | Electrical shock makes cell membranes permeable to DNA. |
| Calcium Chloride/Heat Shock | Chemically-competent cells uptake DNA after heat shock. |
| Incubation on ice | Slows fluid cell membranes. |
| Heat shock | Increases permeability of cell membrane. |
| Nutrient broth incubation | allows beta lactamase expression |
| Arabinose | A sugar that controls the expression of the GFP gene. |
| Transcription Regulation | Process by which cells controls the conversion of DNA into RNA (transcription), thereby regulating gene activity. |
| Plates with arabinose should allow... | the bacteria with the GFP gene to grow and glow green. |
| Plates without arabinose should allow... | bacteria to grow, but not glow green. |
| Gene | segments of DNA that codes for proteins in a lab. Contains instructions to make GFP. |
| Chemical Transformation | Method used to insert plasmid DNA, allows bacteria to take in foreign DNA. |
| pGLO | plasmid used in transformation, carries the GFP gene into bacteria. |
| E coli bacteria | Used in experiments, host cells that receives plasmid. |
| Ultra Violet Light | used to observe bacteria, makes GFP glow bright green. |
| Confirmation Plate | agar plates with antibiotics/arabinose, verify which bacteria were successfully transformed. |
| Inoculation | Placing bacteria onto plates, starts bacteria growth. |
| Supernatant | Liquid used after centrifuge contains dissolved molecules separated from cells. |
| Pellet | solid at bottom after centrifuge, concentrated bacterial cells. |
| TE buffer | Used in DNA work, stabilizes and protects DNA. |
| Lysozyme | Enzyme added to cells, breaks bacteria cell walls. |
| Bacterial Lysis | breaking open bacterial cells, releases DNA and proteins. |
| HIC beads | connecting to binding buffer, beads inside column that binds hydrophobic conditions under high-salt conditions. |
| Equilibration Buffer | prepares the column by setting the right salt/pH conditions before adding sample. |
| Binding Buffer | Helps protein stick to the beads maintaining high salt concentration. |
| Wash buffer | Washes away proteins that are not strongly bound to the beads. |
| Elution Buffer | releases desired protein from the beads by changing salt concentration. |
| SDS-PAGE | Used to analyze and separate proteins by size and shape. |
| Coomassie Blue | dye used in labs to stain proteins so you can see them. Binds to protein and is used after techniques like SDS PAGE. |
| Standard Curve | figure out concentration of an unknown sample by comparing it to known values. |
| Molecular Weight | Helps identify, compare, and analyze molecules like protein and DNA. |
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