| Question | Answer |
| What is the role of TRIS is in the lysis buffer? | Buffer. Prevent pH changes following cell lysis. |
| What is the role of Triton x100 in the lysis buffer? | Non-ionic detergent - disrupts membranes/protein complexes. |
| What is the role of EDTA in the lysis buffer? | To prevent enzyme action by chelating (remove) ions e.g. Mg2+, Ca2+ |
| What is the role of NaCl in the lysis buffer? | Affect ionic strength. |
| What is the role of inhibitors in the lysis buffer? | To prevent proteolysis and other reactions after lysis. |
| What are the types of centrifuge? | Low speed - RCF up to 600g.
Microfuges -RCF up to 10,000g.
High speed - up to 60,000g.
Ultracentrifuges - up to 600,000g. |
| What are the two main types of experimental research? | Hypothesis driven and non-hypothesis driven. |
| What is a negative control? | Control group to which no experimental manipulation is applied. |
| What is a positive control? | Control group which is manipulated in some way to generate the result predicted for the group. |
| What is required in quantitative data presentation? | Title, labels, scales, units, standard deviation/error bars, legends. |
| What is required in qualitative data presentation? | Titles, labels, scales, legends. Images require scale bars. |
| What are the two types of replicate? | Technical replicates and Independent/Biological replicates. |
| What are the types of biomedical research? | Basic, Pharmaceutical and clinical. |
| What molecules are pantent-able? | Small chemical drug molecules.
Naturally occurring biomolecules.
DNA sequences?
Recombinant biomolecules? |
| What is the role of phase I drug trials? | To find:
-The safe dose range
-The side effects
-How the body copes with the drug
-If the treatment shrinks tumours (oncology trials) |
| What is the role of phase II drug trials? | To find:
-If the new treatment works well enough to test in a larger trial
-Which type of cancer it works for (oncology trials)
-More about side effects and how to manage them
-More about the best dose
-Identify any unexpected side effects |
| What is the role of phase III drug trials? | To compare:
-The new treatment with the standard treatment
-Different doses or ways of giving a standard dose |
| What is the role of phase IV drug trials? | To find:
-More info on the side effects and safety
-Long term risks and benefits
-How well the drug works when its used more widely than clinical trials |
| Why do some drug trials fail? | -Tested on patients with advanced disease
-Trial end points may fail to predict overall survival
-Experimental therapy lacks therapeutic efficacy |
| What is a generic drug? | An identical or bio equivalent to a brand name drug in dosage form, strength, route of transmission, quality, performance characteristics and intended use. |
| What are bio-similars? | Molecules similar but not identical to its reference product. Have the potential to cause immunogenic eventsthat are not caused by small molecule products. |
| What is evolutionary medicine? | Any area of medicine related to DNA/RNA changes, adaptation and selection in bacteria/viruses/parasites/humans. |
| What is metabolics? | The study of small molecule metabolic intermediates, hormones and other signalling molecules in individuals. |
| What is the cell lysate? | The supernatant - mainly cytosol, small organelles and solubilised membranes. |
| What happens to the absorbance of Coomassie dye when it is bound to protein? | The absorbance shifts from 465nm to 595nm. |
| What is SDS? | A negatively charged ionic detergent. |
| What do thiol reducing agents do? | They reduce (and break) disulphide bonds. |
| What is the role of SDS in an SDS-PAGE? | It linearises proteins and imparts a negative charge to them. SDS binds strongly to proteins at an approx ratio of 1 SDS per 2 amino acids residues. This allows proteins to be separated due to their resultant negative charge to unit mass ratio. |
| What is the role of Proteinase K? | It is a potent enzyme which digests the proteins present in the Nuclear Pellet. |
| How can you increase the activity of Proteinase K? | Add SDS to the buffer and elevating the temperature. |
| How do you precipitate DNA? | By adding salt and ethanol. |
| How do you rehydrate DNA? | Add a buffer containing EDTA to further inactivate any nucleases still present in the solution. |