Part:BBa_J100184:Experience

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Applications of BBa_J100184

We pipetted 200 microliters of the overnight culture (LB+amp with E. coli) into six different tubes. Two of the tubes contained E. coli with the positive control plasmid J04450, coding for a red fluorescence protein. Two other tubes contained E. coli with the negative control plasmid J100091, which codes for a green fluorescence protein. The final two tubes were filled with E. coli cells containing our modified plasmid with the Phs promoter. Next, we heat-shocked three of the tubes (one of the positive control, the negative control, and the experimental Phs plasmid) in a 43-degree Celsius environment for eight minutes. We allowed the cells to recover at room temperature for thirty minutes, then measured the three heat-shocked tubes and the three control tubes for red fluorescence and absorbance of 590nm wavelength light(to measure density), using a fluorometer and a spectrophotometer, respectively.

Twenty one slots were filled in the spectrophotometer/fluorometer. Three were made from each of the six tubes. Three slots contained only LB and ampicillin as a blank. The light absorbance of the blank was subtracted from the absorbance of each experimental slot. The values on the graph were calculated by dividing the average fluorescence over the average absorbance for each of the six tubes. Error bars indicate standard deviation.

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