In order to perform a DNA extraction from M. guttatus leaves, we began by labeling three 2.0 mL tubes with the sample codes GS1, GS2, GS3. In each tube, three stainless steel beads and a small amount of leaf tissue were placed and attached to the modified reciprocating saw rack to be shaken. The tubes were then spun down in the centrifuge to pull down plant dust. We then added 434 microliters of preheated grind buffer to each tube and incubated it at 65 degrees celsius for 10 minutes in a water bath. We then added 130 microliters of 3M pH 4.7 potassium acetate and inverted the tubes several times and incubated the tubes on ice for 5 minutes. The tubes were once again centrifuged for 20 minutes at maximum power.
Three new sterile 1.5 mL tubes with the sample ID that it was assigned when it was collected were labeled and filled with the supernatant. We then added 1.5 volumes of binding buffer, and 650 microliters of this mixture transferred to Epoch spin column tubes. The column tubes were centrifuged for 10 minutes and the flow through was discarded. The flow centrifuge and flow through steps were completed until the mixture had all gone through. A dry centrifuge with the spin column tubes was then done for 5 minutes. The collection tubes were discarded and and the columns were placed in labeled sterile 1.5 mL microcentrifuge tubes. 100 microliters of preheated pure sterile water to each tube and was left alone to stand for five minutes and then centrifuged for 2 minutes to elute the DNA.