Light spectroscopy has become an incredibly useful tool in analyzing the structure and composition of compounds. Specifically, the principles of absorption, the process by which an electron absorbs radiation to excite from a lower energy state to a higher energy state, and fluorescence, the emission of photons when a material is subjected to radiation, allow for the identification and analysis of unknown concentrations of compounds.1
With that being said, the purpose of this experiment is to spectroscopically assess the concentration of varying DNA and protein solutions. In order to do this, a series dilution will be prepared from bovine serum albumin (BSA) and distilled water. Next, a spectrophotometer, an apparatus that measures the intensity …show more content…
Then, using the same procedure as before, the solutions ' ultraviolet absorptions at 450 and 595 were determined and recorded in Table 4. As for the BCA assay, 100 microliters of each ovalbumin dilution were combined with 900 microliters of BCA, and the resulting solution were analyzed with a spectrophotometer in order to determine the uv absorbances at 560 nanometers (Data displayed in Table …show more content…
As a result, the rule of thumb “1 AU260= 50 micrograms/ ml DNA” is slightly inaccurate. In actuality, about 0.933 AU260= 50 micrograms/ ml DNA. Additionally, since pure DNA has a A260/A280 ratio between 1.8 and 2, and since the experimental ratios of A260/A280 ranged from 1.57 to 1.68, we can determine that the original DNA solution was slightly impure. Lastly, final part of the experiment, the DNA fluorescence assay, yielded an equation of y= 0.0141x + 0.6441 (R2= 0.962) and a molar coefficient of 0.0141
With that being said, the purpose of this experiment is to spectroscopically assess the concentration of varying DNA and protein solutions. In order to do this, a series dilution will be prepared from bovine serum albumin (BSA) and distilled water. Next, a spectrophotometer, an apparatus that measures the intensity …show more content…
Then, using the same procedure as before, the solutions ' ultraviolet absorptions at 450 and 595 were determined and recorded in Table 4. As for the BCA assay, 100 microliters of each ovalbumin dilution were combined with 900 microliters of BCA, and the resulting solution were analyzed with a spectrophotometer in order to determine the uv absorbances at 560 nanometers (Data displayed in Table …show more content…
As a result, the rule of thumb “1 AU260= 50 micrograms/ ml DNA” is slightly inaccurate. In actuality, about 0.933 AU260= 50 micrograms/ ml DNA. Additionally, since pure DNA has a A260/A280 ratio between 1.8 and 2, and since the experimental ratios of A260/A280 ranged from 1.57 to 1.68, we can determine that the original DNA solution was slightly impure. Lastly, final part of the experiment, the DNA fluorescence assay, yielded an equation of y= 0.0141x + 0.6441 (R2= 0.962) and a molar coefficient of 0.0141