Beta Galactosidase Essay

From this graph and chart we can see that the higher the concentration the higher the absorbance, all the different concentrations were tested at the same wavelength (625nm). Also we can determine our unknown substances concentration by using the absorbance we got for it. The red dot on the graph followed by the line towards the horizontal axis indicates that the concentration of fast green was 34% or 5.1x10-3.

10 microliters of the sample is then added and the assay absorption is measured at 340nm. If absorbance was above 1.5, samples were diluted.

The ending result of this experiment confirms that as five test tubes are lined up with the varying level of absorbance, different results in the level of absorbance will appear as well, this is visible in above table. Thus, this is due to the varying amount of water in the solution. The blank sample had a 0.30 in its level of absorbance.

3. State the name and structure of the functional group for each type of biologically

The Beers Law calibration experiment used many concentrations of crystal violet solutions. Each of these solutions were test and analyzed in order to determine the absorbance of each concentration The results were than graphed and produced a slope of 1.00E05 with an intercept of -2.21E-02.

The dark, navy blue colored graph represented the absorbance curve for the S1 sample. The red colored graph represented the absorbance curve for the S2 sample. The green colored graph represented the absorbance curve for the P1 sample. The purple colored graph represented the absorbance curve for the P2 sample. The gaps between the P2 curve was due to the oversaturation that led to the inconclusive spectrophotometer readings. The blue colored graph represented the absorbance curve for the P1 low salt sample. The orange colored graph represented the absorbance curve for the P2 low salt sample. The light blue colored graph represented the absorbance curve for the P1 medium salt sample. The light pink colored graph represented the absorbance curve for the P2 medium salt sample. The light green colored graph represented the absorbance curve for the P1 high salt sample. The light purple colored graph represented the absorbance curve for the P2 high salt

8. If an enzyme is present, it also lowers the activation energy needed to get the reaction started.

The EMG 26 strain cultured on the sixth and seventh lane is a competent producer of _lacI_ gene. This gene produces _lac_

After the substrate solution was added, five drops of the enzyme were quickly placed in tubes 3, 4 and 5. There were no drops of enzyme added in tubes 1 and 2 and in tube 6 ten drops were added. Once the enzyme solution has been added the tubes were then left to incubate for ten minutes and after five drops of DNSA solution were added to tubes 1 to 6. The tubes were then placed in a hot block at 80-90oC for five minutes. They were then taken out after the five minute period and using a 5 ml pipette, 5 ml of distilled water were added to the 6 tubes and mixed by inversion. Once everything was complete the 6 tubes were then taken to the Milton Roy Company Spectronic 21 and the absorbance of each tube was tested.

The absorbance is measured using a Plate reader and a Standard curve is generated. Also, the different types of pipetting techniques are assessed in this Assay.

At 0 minutes, tube 2 had an absorbance of 0.175 because it only contained the clear catechol substrate. Tube 3 had an absorbance of 0.775 because the catechol oxidase enzyme made the solution darker. Tube 4 had an absorbance value of 1.081 because this tube contains both the catechol substrate and the catechol oxidase enzyme, which react to form benzoquinone. The same was true for tubes 5 through 7. Tube 5’s absorbance was 1.065, tube 6’s absorbance was 1.078, and tube 7’s was 1.059. At 10 minutes, Tube 2 had a decrease in absorbance to 0.106 due to little to no reaction occurring. Tube 3 had an absorbance of .955 because it also had very little reaction occurring. Tube 4’s absorbance was 2.110 due the reaction between the catechol substrate and the catechol oxidase enzyme, which was turning the solution brown. Tube 5 had an absorbance of 1.645 because the enzyme substrate reaction was also taking place, but at a slower rate than tube 4 due to the lower temperature in tube 5. The absorbance in tube 6 was 1.833, which means that the reaction at this temperature was occurring faster than tube 5, but slower than tube 4. Tube 7 had an absorbance of 1.155 because the high temperature was denaturing the enzyme causing the reaction to slow or even stop. At 20 minutes, the absorbance of tube 2 stayed about the same with 0.091. Tube 3 had an absorbance of .902, which did not change much

The absorbance with SDS shows a rapid increase, before the graph begins to increase more slowly and uniformly. The absorbance levels off at 0.9nm. The absorbance with no SDS remains at a constant level of 0.01nm for the entire experiment. The absorbance of the control solution is also constant throughout the experiment. This shows the absorbance of the compounds in the solution without the ovalbumin, by taking this figure away from the other recordings, it is possible to discover the absorbance for ovalbumin alone.

An enzyme is a catalyst. Catalysts are known for speeding up the rate of reactions by lowering the activation energy of the biochemical reaction. (Reece et al., 2011)

The goal of the lab was to discover the concentration of phosphate in various colas in order to help with a project for a major cola company. Before we could accomplish this final goal, a Spec 20 was used to test three colors of food colorings. A Spec 20 is an instrument used to “measure the intensity of a light beam before and after it passes through a sample and compares these two intensities” (What does a Spectronic 20 measure). The instrument is able to measure the percent transmittance and absorbance of a given sample. It works by taking light of various wavelengths and passing them through a monochromator. A monochromator is a device that transmits a wavelength of light. The machine diffracts the wavelengths. The sample that is placed in a small container, called a cuvette, absorbs various wavelengths. This depends on the particular sampled being tested. The amount of light absorbed is projected by the machine on a screen, in our case, a digital screen (How Does a Spectronic 20 work?). By doing the lab with the food colorings, we were able to properly learn how to use the Spec 20 and also learn the relationship between absorbance and transmittance. Percent transmittance is the amount of light entering the sample in the cuvette divided by the amount of light leaving the sample, multiplied by 100. This percent can then be converted to absorbance, which is a measure of the capacity of a substance to absorb light of a specified wavelength (Absorbance). It is converted by

We have timed each of the three experiments for 5 minutes to make sure that our results are correct. Thanks to the program, we got the transmittance and the absorbance, together with the graph portraying our results.