Molecular genetics lab
Experiment 1: Isolation and analysis of DNA
Present the standard curve based on the given data, and indicate on the graph your own absorbance values. Have informative titles, correct labels, and a brief description of your results that allows for easy interpretation. (4 marks) (this should be a scatter plot)
Calculate the DNA yield from your initial blood sample in µg/ml. Explain your calculations. (4 marks)
How do your results compare with data published in literature? (2 peer reviewed journal or product manual references are required. References have to be about DNA extraction from mammalian blood, not necessarily equine. Use Harvard style in-text references, and see Q8 for list of references.) (4 marks)
Experiment 2: Analysis of phosphate acquisition mutants of E. coli.
Present the table from the handout on which you recorded your results. (No marks, will be used as a guide for marking Q2)
Strain Growth on Pi
plates (yes/no) Growth on G3P plates (yes/no) AP-ase
assay colour
1 No Yes Yellow
2 No Yes No change
3 Yes Yes Yellow
4 Yes Yes No change
Use your results to determine the genotype of each strain (bacteria are haploid, so you would use the terms “(loss of function) mutant” or “wild type”) and complete the table below. Explain how you reached your conclusions for the genotype of each strain, including the relevance of controls (4 marks)
Genotype
Strain Alkaline phosphatase Pi transporter
1
2
3
4
Suppose the DNA sequences for the alkaline phosphatase and the Pi transporter genes from strain 1 were determined. If both genes are found to contain a single base change compared to the wild type sequences, what could you conclude about the effect of each mutation on the encoded protein? (4 marks)
Based on two scientific journal articles, briefly describe two loss of function mutations of Pi transporter gene(s) in Escherichia coli. (Use Harvard style in-text references, and see Q8 for list of references.) (4 marks)
Write a list of references in Harvard style for your references in Q3 and Q7 (6 marks)
