This project is part of Marin Academy's week long course "Inside out of Fungi" that took place February 22nd-26th which looked to expose students to the diversity of fungi around them through many different lenses including habitat, macromorphology, microscopic-morphology, cellobiase activity, molecular biology through DNA sequencing and ultimately phylogeny mixed with a bit of taxonomy.

Below is the schedule for the week:

Monday
1)Hike at Phoenix Lake park to find and collect fungi (permit obtained through the Mt.Tam watershed).
2)DNA extraction from fungi and PCR amplification using Bio-Rad's Fish DNA Barcoding kit but with M13 flanked ITS primers
3) Set up spore prints

Tuesday
1) Built gel electrophoresis boxes using Bio-Rad's STEM Electrophoresis kit and separated Skittle's candy dyes based upon their size and charge.
2) Run QC gel of yesterday's PCR fragments and send good bands to Sequetech for sequencing.
3) Do crude cellular extractions from fungi and test the various species for cellobiase activity at room temp using Bio-Rad's Biofuel Enzyme kit.

Wednesday
1)Hike out to Dominican Campus looking for and collecting more fungi; urban environment to oak lands.
2)test cellobiase activity of fungi at elevated temperatures
3)Talk on how mushrooms are identified and what is a species?

Thursday
1) Use microscopes to look at small features of fungi (spores, basidium, etc.
2)Phylogeny of mushrooms found so far.

Friday
1) Work through sequences received back from Sequetech in googles code's SeqTrace. Make alignments of the FW and RV traces and trim for high quality sequence.
2)Update iNaturalist observations with ITS sequence and sample number that relates back to gel image
3)Read article "The shape of fungal ecology: does spore morphology give clues to a species' niche?" Discuss experiments from article, experimental design, phylogenetic constraint and life cycles.

Sequenced Fungi can be found in this list
http://www.inaturalist.org/observations/2718379
http://www.inaturalist.org/observations/2707529
http://www.inaturalist.org/observations/2712426
http://www.inaturalist.org/observations/2708702
http://www.inaturalist.org/observations/2707580
http://www.inaturalist.org/observations/2709074
http://www.inaturalist.org/observations/2709065
http://www.inaturalist.org/observations/2708886
http://www.inaturalist.org/observations/2707562
http://www.inaturalist.org/observations/2718307
http://www.inaturalist.org/observations/2709142
http://www.inaturalist.org/observations/2707570
http://www.inaturalist.org/observations/2708892
http://www.inaturalist.org/observations/2707551 less ↑