We‘re Not Yolking Around! – Week 4 Update

Okie dokie. Well, this week we decided to go out on a limb to see what direction to take our project. You may (probably not) remember that I ended my post last week with the idea that we are looking to incorporate another age of embryo into our project, either much younger or much older, in order to have a better chance of getting interesting and more exciting results from our RNA extraction. Dissecting older embryos is probably the more reliable/easier option since we would not have to change our protocol all too much…but what fun is choosing the easier route? Thus, this week we embarked on attempting to extract RNA from younger, almost non-existent, embryos.

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A Success in Trials

After dealing with some plasmid troubles for a few weeks, I’ve finally found some success! After changing bacterial types, plasmid preparation kits, and eventually the DNA that I was transforming from, I was able to get a decent yield. In the end, I had to use DNA from a different tube, one that I had already prepped during the school year. I was originally trying to transform from our original tube, but I have a feeling that the concentration in that tube was just too low for the plasmid to copy well in bacteria. During the plasmid prep that I got to work, I still got a fairly low yield, but it is certainly workable. Luckily, I only need to transfect in about 200 ng of plasmid per well, so as long as my DNA is at least 100 ng I can use it.

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Week 4

After meeting with my advisor last week, I was encouraged to add a new element to my project, to strengthen the relevance of the data I am currently collecting. This added element is a sub-project, which would provide more detailed information on how the bacteria is resistant to ethanol. This sub-project consists of growing the same strains of bacteria in a minimal medium that contains just a low concentration of ethanol, but in which that low concentration of ethanol is the only source of carbon. This experiment would show whether the bacteria is simply resistant to the ethanol, or whether in the process of acquiring resistance, it has also acquired the ability to metabolize ethanol. Since the process of making minimal media requires practice, one of the experiments I did this week was using M9 minimal media without ethanol, with a carbon source that is known to be effective, and practicing the procedure with this practice-M9 media. I also continued the experiment that I had been working on, re-doing the readings for MJ1 and VF7744 at 4.0%, 4.5%, 4.6% and 4.7%. I also tried to perform the full experiment on at least one more strain, however, my strains did not grow during the 3 hour subculture, perhaps because the original plates I had used to innoculate them were old, so I had to scrap this week’s new experiments and will start new strains with newly streaked plates next week.

Week 3

This week I continued my work on figuring out the degree of resistance of V. fischeri strains to each percentage of ethanol. This week I worked primarily on the strains VF7744 and MJ1. I performed the same procedure as before, but with these new strains, at each of the concentrations of ethanol between 4.0% and 4.9%. Though I ran the experiment for all percentages of ethanol at the same time, I encountered a similar problem to the one I had last week, where the 4.5%, 4.6 %  and 4.7% strains had unusually high growth in comparison to both 4.4% and 4.8%. After further troubleshooting, I discovered that the 4.5%, 4.6% and 4.7% stock media had become contaminated, but since it was being kept in the fridge the contaminant wouldn’t grow until the experiment was taking place, showing up as exceedingly high growth in comparison to the blanks when tested in the spectrophotometer. I tested this by allowing the 4.5%, 4.6% and 4.7% stock solution to grow for 24 hours in a shaking incubator, without being innoculated, and found that it was indeed contaminated.  This week, I had to troubleshoot a problem with the 4.0% ethanol strains, in which the tubes innoculated with 4.0% ethanol did not seem to have much growth, whereas the tubes with higher concentrations did. The most likely cause of this is that it was contaminated with something that would prevent growth (e.g. antibacterial soap) during use at one point.  To solve this problem, next week’s experiments will include a redo of both MJ1 and VF7744 at these concentrations, with newly made media.

Week 2 at the National Assembly for Wales

I can’t believe I’ve already finished my second week! It’s gone by so fast I’m doing my best to enjoy every moment. I’ve spent all week working away at my long-term project on homelessness. The first part of my research is about rough sleeping – homeless people who are forced to sleep on the streets. I’ve been looking into services that are provided to help these people off the street, as well as into the best housing options to permanently get them out of homelessness. Housing First, the program of taking someone off the street as soon as they get there and putting them in permanent housing coupled with all the necessary services to keep them there, has gained international traction and seems to be slowly taking hold in the UK. It’s incredibly interesting to find out about what countries are doing to help their homeless populations – through Housing First, Finland has nearly eradicated rough sleeping! A common argument is that the program is too expensive but several studies have proven that over time, the cost of putting someone in a Housing First program is nearly half of what it would cost to leave them on the street.

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