#Gel Electrophoresis
- Buffer (TBE)
- Restriction Enzyme- cuts DNA at a specific restriction site which is often palindromic. This is because enzymes are often dimers and are oppisite mirror images of each other. They are often used in genetic engineering and modification.
- Agarose-the material normally used for the 'gel' in gel electrophoresis
- Loading Dye
- Power Source
- Electrostatic forces- DNA is negatively charged so it will go towards the positive ("runs towards red').
- DNA ladder (Molecular weight marker)
- CRISPR Cas 9- this method of gene editing is replacing restriction enzymes because it is easier and more efficient.
- Resolution
- Stock solutions- solution kept at a high concentration in order to be diluted down for use
- Positive, Negative, Sensitivity Controls
- Error and uncertainty
Gel electrophoresis is one of the most basic techniques in the molecular biology laboratory. This week's lab will give us a new skill, refresh some old skills, and introduce the idea of designing a 'scientific' experiment.
How much of the following will we need in order to make 250ml 5X TBE given the following recipe?
| Reagent | F.W. or starting conc. | final conc. |
|---|---|---|
| Tris | 121.4 g/m | 445mM* |
| Boric Acid | 61.83 | 445mM |
| EDTA | 0.5 M | 10mM |
| *milli-Molar |
Hint: In each case, your final solution volume is 250ml - calculate each reagent independently.