ellie9922

Okay, so I'm trying to work out the effect of substrate concentration on reaction velocity. I did an assay, using PNPG in varying concentrations. I then need to work out the reaction rates in order to graphically determine the Km and Vmax for each reaction. So, my results are: (volume of PNPG & absorbance reading) 0.1 (PNPG) = 0.078 (absorbance) 0.2 = 0.124 0.3 = 0.268 0.4 = 0.315 0.6 = 0.507 0.8 = 0.632 1.2 = 0.724 To convert into concentration, I used the Beer-Lambert law. I had already calculated the conversion factor, of 333.3, from a previous experiment. This gave me concentrations of: 25.997 79.992 89.324 104.990 168.983 210.646 241.309 I need to know how to convert this into initial reaction velocity in nanomoles per minute. I believe, as I used 10ml and incubated the solutions for 5 minutes each, I have to times by 10 and divide by 5. So, for the first tube: 26 x 10 = 260 and /5 = 52 I would really appreciate it if somebody told me if i was right or not. As when I attempt to plot graphs using this data, for example an eadie-hoftsee plot, it just isn't looking right. As I have to plot [ S]/v against [ S]. So, every x value on the graph would therefore be 2. Which cannot be right? as [ S] = 26 and v/[ S] = 52/26 = 2