Hannah Holland-Moritz edited Materials and Methods.tex  almost 9 years ago

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All six plants were collected from \textit{Zostera marina} meadows in Bodega Bay, CA. We collected the first plant in [time/date] of 2014. The other five plants were collected in early February of 2015 as a follow-up study of the first. We stored all plants at -80 degrees prior to DNA extraction.   \subsubsection{\textbf{Sample Preparation}}  Plant 1:  Before extraction, each plant was cut into roughly 50 pieces. The first plant was cut into forty-six pieces (summarized in the figure below). Previous data (unpublished) had shown obvious differences between tissue types, so our study focused on fine-scale community variation within each tissue type rather than among them. Briefly, leaves were cut into twenty roughly equally-sized pieces (about five centimeters each). The relative position of each piece was recorded. Similarly, the rhizome was sectioned into six segments. Each rhizome segment was between 1.5 and 2 centimeters in length and contained no more than a single root bundle.  Before extraction, each plant was cut into roughly 50 pieces. The first plant was cut into forty-six pieces (summarized in Because we were unsure of  the figure below). Previous data (unpublished) had shown obvious differences between tissue types, so our study focused on fine-scale community variation within each appropriate amount of  tissue type rather than among them. Briefly, leaves for extraction, half of the roots  were cut sectioned  into twenty roughly equally-sized four smaller  pieces (about five centimeters each). The relative position of each piece was recorded. Similarly, while the other half were left whole. Of the whole roots, three were taken from a root bundle near the shoot, while the other three were taken from a root bundle farther out on  the rhizome was and pooled together into one sample. The  sectioned roots were divided  into six segments. Each rhizome segment was between 1.5 and 2 centimeters in length four pieces we refer to as tip, mid1, mid2,  and contained no more than base. Similar to the whole-root samples three roots were taken from  asingle  root bundle. bundle near the plant shoot while the other three were taken from farther down on the rhizome and pooled according to plant part.  Because we After sectioning, pieces from plant 1 were placed in Zymo Expedition Stabilization Buffer (citation) for storage until extraction.  Plant 2-6:  For plant 2-6 a more targeted approach was taken. For each plant, a single leaf shoot was chosen and divided into roughly equally-sized pieces. Rhizomes  were unsure also divided into roughly equally sized pieces (again each piece contained no more than one root bundle). In the case  of both the leaf and rhizome distance from  the appropriate amount base  of tissue the shoot was recorded. Knowing that segmented roots would yield sufficient DNA  for extraction, half of the six  roots were sectioned selected from each plant for segmentation. Three of these roots came from a root bundle closest to the leaf and three came from a root bundle farther out on the rhizome. As in plant 1, each root was divided  into four smaller pieces while the other half equally-sized segments which we refer to as tip, mid1, mid2 and base.   After sectioning, samples from plants 2-6  were left whole. placed directly into MoBio Powersoil bead-beating tubes and frozen at -20 degrees prior to extraction.