author2
2Instructables1,886Views4CommentsJoined November 23rd, 2016
transdisciplinary artist / researcher in Baltimore MD

Achievements

  • Becoming With Schizophyllum Commune: Create a Sterile Culture From Found Mushrooms

    Glad to help :) so cool that you all were able to replicate the experiments. Let me know if you see any results with the methyl blue as an indicator.

    View Instructable »
  • Becoming With Schizophyllum Commune: Create a Sterile Culture From Found Mushrooms

    If you really want a living specimen you could try to find a mushroom farmer locally, or you can order liquid cultures or petri dishes fairly cheaply online. Mycosymbiotics is a great trustworthy grower. They're in Pennsylvania and sell liquid cultures on Etsy for just 10 dollars each ~ they don't have S.commune, but they do have have over 20 other species to choose from. One of the oyster varieties would be great for dye decolorization, I had good success with pearl oyster. From what I've read all the oyster varieties have pretty high lignocellulosic enzyme production/secretion. Here's a link to the Mycosymbiotics Etsy store https://www.etsy.com/listing/234827841/liquid-culture-mycelium-syringe-your?ref=shop_home_active_3Before doing PCR analysis of fungi, you've gotta extract their DN...

    see more »

    If you really want a living specimen you could try to find a mushroom farmer locally, or you can order liquid cultures or petri dishes fairly cheaply online. Mycosymbiotics is a great trustworthy grower. They're in Pennsylvania and sell liquid cultures on Etsy for just 10 dollars each ~ they don't have S.commune, but they do have have over 20 other species to choose from. One of the oyster varieties would be great for dye decolorization, I had good success with pearl oyster. From what I've read all the oyster varieties have pretty high lignocellulosic enzyme production/secretion. Here's a link to the Mycosymbiotics Etsy store https://www.etsy.com/listing/234827841/liquid-culture-mycelium-syringe-your?ref=shop_home_active_3Before doing PCR analysis of fungi, you've gotta extract their DNA. I found this protocol for a simple microwave extraction from mycelium ~ haven't used it yet but it looks promising. My only thought with the dried mushroom is that if it's super tough and dried out the protocol might not work as well as with tender, living mycelial mass. Here's a doc that I put together summarizing protocols for: DNA extraction, PCR amplification of of a DNA region called ITS region from fungi that is unique in different species, and performing a restriction digest for "DNA fingerprinting" with electrophoresis. If you don't have the equipment to create an agarose gel and do electrophoresis then you can skip the restriction digest and look for a local lab that will do DNA sequencing for you. Usually local universities will have labs that offer sequencing services very affordably. In Baltimore I usually outsource sequencing to a lab at Johns Hopkins University and it's only 6 dollars per reaction, so pretty affordable. https://docs.google.com/document/d/1QXOMzBIf8EbQDCFGZvVUKJN_JQeIdprzRgwQyA3J6aA/edit?usp=sharing

    View Instructable »
  • Becoming With Schizophyllum Commune: Create a Sterile Culture From Found Mushrooms

    Ah, okay. YES screen is good, just so you know - in case you eventually want to go further with this stuff, you can also buy a plasmid to make your own yeast estrogen sensor from addgene: https://www.addgene.org/23062/But this would require you have the facilities to do a yeast transformation (to genetically modify yeast with the plasmid) in you school's lab. Probably too big an undertaking for what you want to do right now.As for the Sigma order - I'm not sure if they sell to educators but it's worth a shot. I just did a quick search for "remazol brilliant blue reactive science supply" and looks like fisher scientific also sells it here https://www.fishersci.com/shop/products/remazol-br... I'd search around and check your options. Not all science supply stores require buyers ...

    see more »

    Ah, okay. YES screen is good, just so you know - in case you eventually want to go further with this stuff, you can also buy a plasmid to make your own yeast estrogen sensor from addgene: https://www.addgene.org/23062/But this would require you have the facilities to do a yeast transformation (to genetically modify yeast with the plasmid) in you school's lab. Probably too big an undertaking for what you want to do right now.As for the Sigma order - I'm not sure if they sell to educators but it's worth a shot. I just did a quick search for "remazol brilliant blue reactive science supply" and looks like fisher scientific also sells it here https://www.fishersci.com/shop/products/remazol-br... I'd search around and check your options. Not all science supply stores require buyers to provide credentials for non-hazardous purchases.And yes! You definitely could use a spectrometer to measure the dye degradation. You could do this by growing your mycelium in liquid culture rather than on a petri dish. In liquid culture, the action of the enzymes on the dye will be the same. Just when it comes time to load a sample into the spec, you'll probably want to centrifuge a sample and only put the supernatant into the cuvette so that the mycelium does not interfere with the optical reading. I had good results growing liquid cultures in baffled flasks at 25C shaking on an orbital shaker at 100 RPM, but have seen studies suggest 30C as the temperature for optimal growth https://www.ncbi.nlm.nih.gov/pmc/articles/PMC37415...I didn't isolate the enzymes, this is a more complex task. I simply grew the mushrooms on agar solidified media containing the dye. As the mycelium grew outward from the center of the petri dish, a halo of decolorization grew in those species that were high producers of certain lignocellulosic enzymes.

    View Instructable »
    • Becoming With Schizophyllum Commune: Create a Sterile Culture From Found Mushrooms
      513 views
      9 favorites
      1 comments
  • RyanH291's instructable Open Works Fob Bracelet's weekly stats: 1 year ago
    • Open Works Fob Bracelet
      637 views
      6 favorites
      1 comments