It has been 1.5 weeks since work began. Finally, the plan is somewhat finalized by supervisors. The rest is up to me. Finally, the heart has caught up with the brain in wanting to work. It sort of took 1 month to achieve this. Unfortunately (or fortunately), in only 1.5 weeks, 2 problems have cropped up.
Should the organism be cultured in mosquito or mammalian cell line? Seniors and articles I read grow the organism in mosquito cell line to simulate normal transmission cycle. I agree with this. Although the genomic material of the organism might not have changed with the use of different cell line, the composition of the envelop might. This might have an effect on membrane fusion or perturbation of the intracellular biomolecular homeostasis. For practice sake, I was taught how to grow up the organism in mammalian cell line. Now the question is, should I use this organism, grown in mammalian cell line to do my experiments. Probably I should grow another batch from mosquito and use those grown from mosquito. The ones from mammalian cell lines, I can use to compare with those grown from mosquito if I have the time.
Problem with medium. I was taught to filter the medium, then add FCS. However, one senior suggested I add FCS, then filter. I did what the senior told me, the filter got stuck due to the large proteins in the FCS. So now, the question is, should I use this medium, or prepare a new one. Since large proteins get stuck in the filter, the final composition of the medium is altered. It could impact on the intracellular biomolecular homeostasis again. Preparing the medium would only take a day. However, checking its sterility would take another one to two days.
So yes. I learnt the skills, though of course not close to ideal yet. These "goof ups" are costing me time, precious precious time... BUT if I used the organism grown in wrong organism, my results may not withstand scrutiny. Then it will waste even MORE time...
