Skunk cabbage samples were taken from Lake Mitchell Swamp, Cadillac, Michigan. Samples were taken from clumps, which were designated as plants that could possibly have come from the same seed or underground vegetative growth and were less than six inches apart. A total of twenty-two clumps were sampled from, in four habitat sites. These sites and clumps can be seen on the map of the study area. The first site, on the edge of the swamp, was drier and dominated by tag-alder. The second site was in the middle of the open wetland so the habitat was much wetter and had both tag-alder and cattails intermixed. Located on higher ground and covered with aspen and grasses, the third site was the driest of the other three. The fourth site was located in the open wetland and surrounded by cattails. The clumps themselves are located from our original clump in site one. The vector and distance were taken from plant one, site one to plant two, site one. The second site was located from plant one, site one and then the clumps (plants) in site two were located off of plant one site two. Site three was then located off of plant one, site two and this procedure was done through site four.

We figured the best DNA samples could be found in the new shoots of the plant, so the top of one shoot from each plant was cut using a razor blade. Each sample was then placed in a zip-lock bag labeled with both site and clump number. The sample bags were then kept on ice in a cooler so that the samples did not deteriorate before we could get them back to the lab.

DNA Extraction

PCR is the acronym for polymerase chain reaction. This is a technique that produces many copies of a particular region of DNA for testing. The PCR method takes a small amount of genetic material and duplicates it enough for one to study. The DNA is placed in a vial along with a primer, a polymerase, and the four bases, adenine, cytosine, guanine, and thymine. The double helix of the DNA will separate and the polymerase enzyme will make a copy of each strand. The four nucleotides will match up with each strand, adenine with thymine, and cytosine with guanine. After heating, cooling, and reheating, the DNA will have been duplicated.

RAPD or random amplified polymorphic DNA is the type of PCR we used. It is a fast, simple, and inexpensive way to amplify unknown DNA.

The PCR amplification was completed as follows:

• A "master mix" of all components needed to perform PCR was added to 3 sets of 21 PCR tubes (including one negative control for each set).
• The "master mix" included sterile water, 10X buffer, dNTPs, the primer, and the Taq polymerase.
• The specific DNA was then added to each tube and water was added to the negative control tubes.

All tubes were placed in the PCR machine and run under normal conditions.

Objectives

Facts about Skunk cabbage

Introduction

Materials and Methods

Results and Discussion

Conclusion