Microsatellite Genotyping of Milkweed:

Milkweed and monarchs populations have been steadily declining due to habitat changes and a whole host of other factors for many years. Conservation efforts include adding more diversity to milkweed populations as lower diversity of milkweed is dangerous because it leads to more susceptibility for milkweed extinction. Analyzing the diversity of milkweed is imperative for replanting efforts and conservation goals of both the monarch and milkweed as milkweed is the most important plant for larval monarchs. My goal for this experiment is to evaluate patches of milkweed and quantify genetic diversity so as to help conservation efforts. I will sample and map the stems of common milkweed in five populations that have been sampled for the past four years to discover how diverse these populations are by using microsatellite markers.  Sampling will occur in 5 populations during the month of June, and the distance between the different plants, ramets, will be recorded on site. Once back in the lab, DNA extraction will occur using a specific protocol. After extraction, Polymerase Chain Reaction (PCR) will be done on the DNA to amplify sections of the DNA sequence using 8 primers developed for this specific species of milkweed, Asclepias syriaca. PCR is done using a MyTaq kit and a thermocline machine. The sections of the DNA amplified are microsatellites in this case. Microsatellites are repeated sections of DNA unique to each plant. However, because milkweed is very clonal, or makes clones of itself, many plants in a patch could be genetically identical, or having the same microsatellite sequence, which means there is very low genetic diversity. Once these sections are amplified, fragment analysis can occur so as to compare the length of microsatellites to evaluate the clonality of milkweed. Additionally with use of the ramet density data, the probability of any plants being related can hopefully be correlated to their distance.