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1. Kolbe reaction is for the synthesis of salicylic acid which can alleviate fever and pain:
Here, 4.70565g of phenol was used as the raw material for the synthesis of salicylic acid. Eventually, 4.83420g of salicylic acid was
acquired. Calculate percentage yield of the product. (30 marks)
2. When performing quantitative PCR (qPCR), absolute quantification is usually accomplished by including artificial templates such
as plasmids, oligonucleotides, or purified PCR products that have been accurately quantified by independent analysis. Standard
curves plotted to known concentrations are then created by performing qPCR on serial dilutions of these templates.
3. The MTT assay is used to measure cellular metabolic activity as an indicator of cell viability, proliferation and cytotoxicity. This
colorimetric assay is based on the reduction of a yellow tetrazolium salt (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide or MTT) to purple formazan crystals by metabolically active cells. The viable cells contain NAD(P)H-dependent
oxidoreductase enzymes which reduce the MTT to formazan. The insoluble formazan crystals are dissolved using a solubilization
solution and the resulting coloured solution is quantified by measuring absorbance at 500-600 nanometers using a multi-well
spectrophotometer. The darker the solution, the greater the number of viable, metabolically active cells.
Here the MTT assay was conducted to measure changes in cell viability of a neuronal cell line after treatment with toxic beta
amyloid proteins. Beta-amyloid proteins play key roles in the pathogenesis of Alzheimer’s disease. The original data of the
experiments are shown as follows:
Based on the raw data (absorbance), please plot the relative cell viability (% of control) with standard deviation. Using
statistical analysis software, conduct a paired t-test to determine if there is a significant difference in cell viability after
beta-amyloid treatment. (40 marks)
