Genomic DNA Extraction
The results from the genomic DNA extraction of Dieffenbachia camille were analyzed using a spectrophotometer at the wavelengths of 260 nanometers and 280 nanometers.  The spectrophotometer’s absorption reading at the 260 nanometer wavelength will be used in Beer’s Law to calculate the concentration of DNA, while the spectrophotometer’s absorption reading at the 280 nanometer wavelength will determine the DNA samples’ purity by using the A260/A280.  These results are shown below in Table 1.

Table 1.  This data table displays the absorption values read using a spectrophotometer at 260 nm and 280 nm and the A260/A280 ratio and DNA concentration values calculated using the absorption values.

Real-Time Polymerase Chain Reaction (PCR) Results Opticon Graph Analysis
The results from the analysis of the data collected from the RT-PCR preformed on the tissue samples taken dieffenbachia Camille. First, the general melting curves were screened for potential contamination or abnormal results. The green tissue samples which had primer set A showed abnormal peaks (figure 2). Using the samples from primer set C, the plate read temperature was determine to be 81 degrees celcius (figure 3). Figure 4 and figure 5 are images of the plate read graph at cycle 11 (approx. 81oc). In the 100ng samples (figure 4) the green tissue had a lower Ct value (table 2). In the 50ng samples (figure 5) the white had a lower Ct value (table 2). Figure 6 illustrates the abnormal peaks presented by one of the controls for primer set C.

Figure 2. - A graph of an RT-PCR melting curve containing four tissue samples from Dieffenbachia Camille as well as a control containing no DNA. Identified by color the major peaks signify melting points of the sample. This graph illustrates the abnormal position of the 100ng and 50ng green tissue samples using primer set A.

Figure 3. – A graph of an RT-PCR melting curve containing four tissue samples from Dieffenbachia Camille as well as a control containing no DNA, all using primer set C.  Identified by color the major peaks signify melting points of the sample. Specifically this graph illustrates the optimal plate read temperature of 81 degrees.

Figure 4. – A plate read of green and white RT-PCR tissue samples at 100ng concentration and one control using primer set C at cycle 11 (81 degrees).  The dotted line sets the threshold at which the Ct values are determined (table 2).

Figure 5.– A plate read of green and white RT-PCR tissue samples at 50ng concentration and one control using primer set C at cycle 11 (81 degrees).  The dotted line sets the threshold at which the Ct values are determined (table 2).

Figure 6 - A graph of an RT-PCR melting curve containing four control samples and one tissue sample from Dieffenbachia Camille, using two primer sets as indicated in the legend. The highlighted brown control using primer set C has an irregular peak far

Table 2- The Concentrations, primer set, and Ct value of the evaluated RT-PCR samples. A lower Ct value denotes a higher amount of PCR product. Refer back to figures 3 and 4 for the graphical representation of each samples amplification.

Real-Time Polymerase Chain Reaction (PCR) Results Agarose Gel Electrophoresis
The results from the real-time PCR for the large subunit of Rubisco gene in Dieffenbachia camille were visually analyzed using agarose gel electrophoresis on a 2% (w/v) agarose gel stained with ethidium bromide.  The stained gel was photographed using a BioRad imager.  This gel’s photo can be seen below in Figure 1.  Six different real-time PCR mixes were run on the agarose gel electrophoresis along with a standard base pair ladder.  A large band of DNA at approximately the same location was observed in the lanes containing the real-time PCR mixes with primer set “C” for the 50 nanograms of DNA from the leaf’s white region and the leaf’s green region.  Another large band of DNA was observed in the lane containing the real-time PCR mix with primer set “A” and 50 nanograms of DNA from the leaf’s green region; however, this large band was not found in the same location as the other two observed bands on the agarose gel.  No large band of DNA was observed in the lane containing the PCR mix with primer set “A” and 50 nanograms of DNA from the leaf’s white region.  Neither control lane containing both primer sets with no DNA showed any DNA bands on the agarose gel.

Figure 1.  Photograph of 2% (w/v) agarose gel stained with ethidium bromide for the results from real-time PCR of the large subunit of Rubisco.  Lane one contained the control of primer set “C” with no DNA.  Lane two contained the control of primer set “A” with no DNA.  Lane three contained the primer set “C” with 50 ng of DNA from the leaf’s white region.  Lane four contained the standard base pair ladder.  Lane five contained the primer set “A” with 50 ng of DNA from the leaf’s white region.  Lane six contained the primer set “C” with 50 ng of DNA from the leaf’s green region.  Lane seven contained the primer set “A” with 50 ng of DNA from the leaf’s green region.

Protein Isolation
The results from the protein isolation samples from Dieffenbachia camille were analyzed for their protein concentrations using BioRad’s DC protein assay with the use of a spectrophotometer.  In order to determine the concentration of this experiment’s, a standard curve of absorbance at 750 nanometers versus protein concentration using known protein concentrations.  The data used for the creation of this standard curve is shown below in Table 3.  The data was then plotted onto a scatterplot using Microsoft Excel to determine the standard curve equation.  This is shown below in Figure 7.  The data from the observed absorbance readings at 750 nanometers for the six protein samples with three for each leaf color region is shown below in Table 4.

Table 3.  This data table displays the known protein concentrations and their observed absorbance values at 750 nm used to determine the standard curve.

Figure 7.  Scatterplot of the known protein concentrations versus their respective absorbance values at 750 nm.  The calculated standard curve equation is shown with its R2 value.

Table 4.  This data table displays each protein sample’s absorbance at 750 nm, protein concentration before and after the dilution factor for the respective sample, and the amount of protein sample that will be needed to use 18 µg of protein for later use involving SDS-PAGE and Western Blotting.

Sodium Dodecyl Sulfate Polyacrylamide Gel Electrophoresis (SDS-PAGE)
The results from the protein isolation samples from Dieffenbachia camille were visually analyzed using SDS-PAGE stained using Coomassie Blue.  Besides the standard protein marker ladder, no protein was visibly seen on the SDS-PAGE after staining with the Comassie Blue.

Figure 8.Commasie blue stain of an SDS PAGE containing two tissue samples of Dieffenbachia protein and a molecular weight ladder. Lane-1 18ng Green tissue, lane-3 18ng White tissue, lane-5 molecular weight. No discernable results from the lanes containing tissue. 

Western Blotting
The results from performing a Western Blot on an unstained SDS-PAGE were visually analyzed for the presence of the large subunit of Rubisco using a primary chicken anti-Rubisco antibody followed by a secondary goat anti-chicken antibody.  The photo of the Western Blot using a BioRad imager is shown below in Figure 9.  The standard protein marker ladder did not show any band for the large subunit of Rubisco.  The protein sample from the leaf’s green region and the protein sample from the leaf’s white region both produced a dark band for the presence of the large subunit of Rubisco at the same location in their respective lanes.

Figure 9.Photograph of Western Blot Immobilon-PDVF Transfer Membrane probed for the large subunit of Rubisco by a primary chicken anti-Rubisco antibody followed by a secondary goat anti-chicken antibody.  Lane five contained the standard protein marker ladder.  Lane one contained the 18ng protein sample from the leaf’s green region.  Lane two contained the 18ng  protein sample from the leaf’s white region.