click below
click below
Normal Size Small Size show me how
Lab and Study skills
key info from module
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. |