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Lab and Study skills

key info from module

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.
Created by: chloesemple
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