Arrival
Everyone that I have come into contact with so far has been incredibly friendly and welcoming, which has helped me settle into the laboratory in a relatively short space of time. My supervisor for this project is a fantastic second year PhD student named Harriet, who has involved me with experimental planning as well as helping me with practical laboratory work. On the first day, I attended a seminar given by Dr. Evan Reid on the role of spastin in membrane trafficking; and how this may contribute to hereditary spastic paraplegias (HSPs). This is an area of research that I have barely touched upon during my undergraduate study; which meant the talk itself was very informative as well as interesting.
Laboratory Work
Experiment 1: Cell Wall Stress Plates
This involved preparing YPD (the media used to support Candida growth) plates which contained various chemical compounds that exerted stress onto particular cell wall components. I then "stamped" Candida onto each plate in a pattern that was comprised of wild-type and mutant strains arranged into decreasing cell concentrations. The apparatus that I used to stamp the Candida was a nifty instrument that Harriet referred to as a frog and hedgehog.
As of now, our plates appear to be doing well and it looks as though we have managed to gain some interesting results!
Experiment 2: Alcian Blue Staining Assay
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| View of Firth Court from the Laboratory |
Everyone that I have come into contact with so far has been incredibly friendly and welcoming, which has helped me settle into the laboratory in a relatively short space of time. My supervisor for this project is a fantastic second year PhD student named Harriet, who has involved me with experimental planning as well as helping me with practical laboratory work. On the first day, I attended a seminar given by Dr. Evan Reid on the role of spastin in membrane trafficking; and how this may contribute to hereditary spastic paraplegias (HSPs). This is an area of research that I have barely touched upon during my undergraduate study; which meant the talk itself was very informative as well as interesting.
Laboratory Work
Experiment 1: Cell Wall Stress Plates
This involved preparing YPD (the media used to support Candida growth) plates which contained various chemical compounds that exerted stress onto particular cell wall components. I then "stamped" Candida onto each plate in a pattern that was comprised of wild-type and mutant strains arranged into decreasing cell concentrations. The apparatus that I used to stamp the Candida was a nifty instrument that Harriet referred to as a frog and hedgehog.
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| Frog and Hedgehog. Used to stamp Candida onto multiple YPD plates. |
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| Control Candida Cell Wall Stress Plate. Each row contains a different candida strain; and each column contains a particular cell concentration. |
As of now, our plates appear to be doing well and it looks as though we have managed to gain some interesting results!
Experiment 2: Alcian Blue Staining Assay
This involved staining mutant and wild-type strains with alcian blue and subsequently conducting an absorption assay. In order to calculate alcian blue concentration from absorbance, we first had to prepare a standard curve from known concentrations of alcian blue. We then obtained absorbance measurements from the supernatant of stained cells, to in turn infer the amount of alcian blue that had actually bound to our samples.
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| Range of Alcian Blue Concentrations used to Create Standard Curve |
We did get some results from this experiment, however as this was some what of a preliminary run through, we didn't include repeats and so can't do any statistical tests on our current data. In addition, after reading up about the assay over the weekend, I think we can do some additional calculations in order to present our data in a way that mirrors that seen in publications. Therefore, I think I will be giving this another go next week and hopefully producing some more meaningful results.
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