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Characterization; team TEL HAI-MIGAL 2019:

Results: In order to examine the expression level of the surCD19 protein on the cell membrane, K562 cells were cultured and then transfected with 10µg mRNA. The expression was confirmed and quantified (% of expression within a population of cells) by FACSCallibur 45 minutes after cells were incubated with an αHuman c-Myc conjugated APC antibody or a HA’ conjugated APC antibody 24hrs post-transfection. The controls for this experiment were irrelevant mRNA and a positive control. The FACS was calibrated according to the irrelevant mRNA transfected cells incubated with the antibody. In Figure 1, the expression of the c-Myc tag was examined and showed a 5.01% background from the irrelevant mRNA, while the positive control showed 93.57% expression. The surCD19 protein showed only 10.16% expression in comparison to the controls. In Figure 2, the irrelevant mRNA showed an expression of 3.75% and our positive control only expressed 8.82% of the HA tag. Similarly, to the expression described by the Team Enable of China, there was a high level of surCD19 expression; our experiment showed 52.78% expression.