The objective of the lab is to be able to determine the proper dilution for the crude lysate, which will yield an O.D. of about 0.1-0.7. It will then be to determine its concentration using the Bradford Method.
Data/Presentation In Table 1, a series of calculations were conducted in order to find the specific activity for Tubes 2-5 using data from the 1:250 dilution. As the molar concentration of the product, PNP, increased with each successive tube the absorbance also increased. The specific activity of the enzyme was highest in Tube 5 where the volume of diluted enzyme was highest. On the other hand, the specific activity of the enzyme was lowest in Tube 2 where the volume of diluted enzyme was also the lowest. Figure 1: A graph of the U/mL for the undiluted enzyme versus the volume of diluted enzymes was created. As the volume of the diluted enzyme increased the U/mL for the undiluted enzyme also increased. The graph steadily increases at first and then it begins to sort of level off resulting in a parabola. In Table 2, the concentrations and the absorbance at 595nm of both the BSA protein standard as well as the crude lysate (for the 1:100, 1:500, & 1:1000 dilutions) were calculated. For the BSA standard, as the concentrations of the solutions increased the absorbance also increased. The absorbance for the 1:100, 1:500, and 1:1000 dilutions were 3.781, 1.717, and 1.321, respectively. The concentrations for each were .104, .032, and .018,
