Since the last time I posted a blog, I have conducted my experiment about 4 or 5 times. Each time, I have improved on my techniques. The hardest technique that I have yet to perfect would be the injection of morpholino oglionucleotides into the zebrafish embryos. This technique is very difficult and is the basis of my experiment. As far as extracting RNA, making cDNA, running PCR and gel electrophoresis, I believe i have those down pact at this point. I have done them enough times to be confident in my abilities.
Because I can’t get data without injecting properly, I haven’t retrieved as much data as I would have liked. However, from the data that I have collected, I found that the morpholino oligonucleotide that I have been trying to inject, has been knocking down the gene I’ve been trying to knock down. My plan is to work throughout winter break to continue practice of injecting the fish embryos. I am confident I will be able to do it properly soon. After I can properly inject my fish embryos, my next step is to kill the hair cells by copper treatment to measure the rate of growth and cell number of the hair cells.