Labwork

Following the collection of data and samples, it was time for lab work to begin. The first step of my procedure was DNA extractions. With around 800 plant samples, I quickly realized financially and time wise doing all 800 would not be practical. This led to many discussions on how to subsample. Was it better to do more transects, with fewer plants from each transect? More plants, with fewer coverage of overall transects? How do you account for the difference in densities between transects? Overtime, with more and more discussions, it became clear that sampling more transects would be a better option, even if that meant fewer plants per transect. Additionally, for any transect with ~30 or fewer plants, the entire transect would be sampled. For any plant over that, a subsample would be done using a random number generator to randomly select which plants should be extracted.

Though all ~800 have plant data collected for them from the field, in the end only around 400 will be looked at intensively. However, the benefit of doing it this way is that if in the future I would decide to expand my experiment, all sampling has already occurred so it would not be hard to add on to it.

Once I figured out the subsample, I began doing extractions. Extractions are done using liquid nitrogen, grinding up the plant, adding a buffer, and heating the sample to denature the DNA. This process takes a fairly long time. It is very important to sterilize everything in between too, because any bit of contamination could ruin the results. 400 plants later, and the extraction process was finished by mid July. Up next, Polymerase Chain Reaction (PCR)!!

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