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Abstract   Introduction    Materials & Methods    Results    Discussion    Literature cited

All the procedures for this experiment can be found on the James Madison University Biology 480/580 website.

Three leaf fractions were selected for this experiment, green leaves, white leaves and the green leaf margins of mostly white leaves of A. schefflera. These three plant tissues were processed to isolate the protein and determine the protein concentration.

A quantity of 120 µg of the protein isolated from each plant tissue was loaded into two polyacrilamide gels. One gel was used to visualize the proteins isolated by staining with comassie blue and one gel was used in a western blot. The western blot membrane was probed for the RuBisCO large subunit with a chicken antibody and a secondary goat anti-chicken antibody and the alternative procedure for a light reaction was utilized with a five minute exposure.

        DNA from the three plant tissues was isolated and the concentrations were determined using a spectrophotometer. To quantitiate the number of copies of the  RuBisCO large subunit gene in each leaf fraction a real time PCR was performed using the rbcl2f primer set. A standard curve was also constructed using a plasmid containing the RBCL gene, and an agarose gel of the real time PCR products was performed to ensure that the correct segment of DNA was amplified.