Results

Protein Isolation

After isolating protein, a DC protein assay was performed to determine the concentration of protein in our samples.  A standard curve (Figure 1) was created with samples of known protein concentration (Table 1).  The standard curve was produced using a Spectrophotometer at 750nm. The linear trend line had an R² value of 0.9299. There was a point that was removed from the data before plotting, and it was the 1.5 mg/mL standard.  The equation which was derived from the linear relationship helped to determine the protein concentration in our samples (Table 2) along with  the volume necessary to run the Western Blot.  The flowers, no water sample had the greatest amount of protein present (19.08 µg/µL) while the leaves, water sample had the least amount of protein present (14.8 µg/µL).

Western Blot

The samples were run on an acrylamide gel (Figure 2) and then transferred onto a membrane using electrophoresis. After the membrane was incubated with the two antibodies, and exposed with the Chemiluminescent camera, it was observed that only the molecular weight marker was transferred to the membrane.

DNA Isolation

After DNA isolation, the concentration and purity of our samples was determined using the Nanodrop (Table 3).  All four samples contained similar DNA concentrations (Table 3).  The DNA purity of the flowers, water sample was much higher than the purities for the other three samples (Table 3).  Real-time PCR was performed to amplify the RBCL gene (Figure 3).  Three samples had similar C _t values (Figure 3).  These samples were leaves, water; leaves, no water; and flowers, no water.  The melting temperature of the samples was also quantified during real-time PCR (Figure 4 and Table 5).  All of the samples including the negative control (no DNA) had similar melting points (Figure 4).  The samples were then analyzed on an agarose gel (Figure 5).  PCR products were only observed in two of the samples:  leaves, no water; and flowers, no water (Figure 5).  PCR products were also observed for the molecular weight marker as well as the negative control (Figure 5).  The real-time amplification of several standards containing known amounts of Rubisco DNA was also performed (Figure 6 and Table 6).

Image from: http://www.mooseyscountrygarden.com/perennial-plants/chrysanthemums.jpg. Accessed 2 Oct 2007