Which mutation/cell line looked the most promising at 6% in the last assay you ran? Have you run a full IL3 withdrawal assay for all 4 lines of cells? Is there a reason to expect that all 4 mutations would behave similarly at 6%?
Thank you Veronica for the great questions. They make me dive deeper into the reasoning behind my research and also prompt me to be more specific when I describe a certain experiment.
So I've put a decent amount of work in over this break. Still, my paper is far from completion. I have the background and procedure sections in good places, though I still need to change my procedure from commands to past tense. I have included a picture of my western blot and would like to know where that would go. My largest worry right now is having enough content, after talking to Kristin, she said her paper is roughly 19 pages in length right now.
Yesterday, I went to the lab to start a new western blot.
With the same original Ba/F3 cells that were used for this IL3 withdrawal assay (my mentor thawed some cells from vials that I froze down before, so now there are a lot of healthy, fresh Ba/F3 cells in the incubator), I'm working on this next western blot. My mentor helped me lyse the cells with lysis buffer before I came, so that the cell lysate would be ready for me to begin the gel right after school.
I went to the lab on Monday to begin a new IL3 withdrawal assay with 6% serum. This assay includes all four mutations from the four different cancers (3 in NTRK1 and 1 in NTRK2), as well as NTRK1 and NTKR2 WT, and the control, pMSCVpuro (retroviral vector w/no NTRK). Just for more info: pMSCVpuro is a retroviral vector (plasmid) "optimized for expression of a gene in hematopoietic, embryonic stem, or embryonic carcinoma cells" (AddGene).
I re-read the NWSE/ISEF guidelines and I am wondering if I should be filling out continuation paperwork. If so, I could do that January 6-7th.
The thing is, I am not using any information from last year's project (new material PbS instead of last year's PbSe, totally new algorithms and model, multijunction aspect completely revises my procedure), so this is not a continuation project.
Each algorithm was implemented in JAVA programs. First, raw NASA solar spectrum data was read into the program as a XLSX file and data-mapped into a TreeMap, an ordered organizational structure in JAVA. The Solar Spectrum and Photon Flux algorithms were implemented and programmed to prepare the data for future computation. Then, absorption spectra, quantum mechanically cloud-computed beforehand for each quantum dot diameter (1 nanometer to 5 nanometers, in increments of 0.5 nanometers), were read into the program as TXT files and data-mapped into a separate TreeMap.