GMU:DIY Biolab “Driver’s License”/Jan Georg Glöckner: Difference between revisions

Latest revision as of 15:56, 31 March 2018

The digital lab-book ( this is the unedited version of the lab-book, that will be the most up-to-date document)

supermarket piracy programm

Oyster mushroom cloning series stage 1 (obtained from Globus supermarket; polish origin)

Brown Button mushroom cloning series stage 1,2 and stage 3 [petri dishes A to D and G.H not shown] ( obtained from Rewe supermarket; german origin)

Artistic Abstract

To address the growing issue of patented living organisms, I pirate the genome of store-bought mushroom-products, by reverse engineering the product to its viable state of mycelium. I then distribute the organism, for everyone to culture and distribute, in a respectful way and to form mutual diplomatic relations. As the genetic makeup of the strains change through cultivation, the creative commons licence is applied to the genome to keep it from being obscured by companies.

CC BY-SA 4.0

Scientific Abstract

Specimen of mushrooms obtained from supermarkets are cloned and the developing strain is purified, than stored in a cold-storage strain-bank. Via reverse-engineering the strain from a commercial product is extracted and cultured. The strains will be given away, and kept available.

Method

For every attempt of cloning at least seven petri-dishes filled with PDA- medium are inoculated. The scalpel is sterilized in a hot flame till it glows red. It then is cooled in the receiving petri dish. The mushroom is ripped apart in the middle. Tissue then is taken from just above the gills, inside the stem or just under the cap. It is inserted in the hole that the hot scalpel made. The dishes are labeled and incubated in the incubator (23°-25° C). The mycelium that emerges is purified by running it through subsequent petri dishes till there is no visible evidence of contaminants. Then it is stored in test-tubes filled with PDA- medium at temperatures below 5°C. Two tubes per strain, enveloped in a zip-loc-bag. ( liquid suspended-animation technique will be introduced.)

References

Paul Stamets and J.S. Chilton: The Mushroom Cultivator; Sterile Technique and Agar Culture: Starting a Culture From Live Tissue (p. 29) 1983 Agarikon Press Olympia Washington ISBN: 978-0-9610798-0-2
Paul Stamets: Growing Gourmet and Medicinal Mushrooms (Third Edition); Culturing Mushroom Mycelium on Agar Media: Starting a Mushroom Strain by Cloning (p. 88); 2000 Ten Speed Press ( Random House New York) ISBN: 978-1-58008-175-7

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-'''The digital lab-book'''
+[[/The digital lab-book|The digital lab-book]] ( this is the unedited version of the lab-book, that will be the most up-to-date document)
-'''25. October'''
-inoculated cooked cardboard with Pleurotus pegs.
+[[/General methods|General methods]]
-'''31. October'''
-starting to grow vibrio phosphoreum from a batch that was labelled Photobacterium phosp. and seemed to be inoculated somewhere in 2016. (the writing on the petri-dish was rendered unreadable)
+==supermarket piracy programm==
-cooked 100ml of LA-Medium and left it overnight in the fridge.
+Oyster mushroom cloning series stage 1 (obtained from Globus supermarket; polish origin)
+<gallery>
+File:Pleur.ostr.SAD1.JPG|
+File:Pleur.ostr.SAD1 alternateview.JPG|
+File:Pleur.ostr.SAD2.JPG|
+File:Pleur.ostr.SAD3.JPG|
+File:Pleur.ostr.SAD4.JPG|
+File:Pleur.ostr.SBD1.JPG|
+File:Pleur.ostr.SBD2.JPG|
+File:Pleur.ostr.SBD3.JPG|
+File:Pleur.ostr.SBD3 alternateview.JPG|
+</gallery>
-''on plastic petri-dishes: I decided to use only glass-petri-dishes, as they give me the opportunity to sterilise medium inside the dishes, thus limiting to chances for contamination  greatly. and they produce less waste, are to be recycled if broken, if not broken have a unlimited shelf life and usage time.''
+Brown Button mushroom cloning series stage 1,2 and stage 3 [petri dishes A to D and G.H not shown] ( obtained from Rewe supermarket; german origin)
-migrated the pleurotus from the plastic mushroom supermarket dish to a tupperware. no sign of growth.
+<gallery>
+File:button E1 alternateview.JPG
+File:button E2.JPG
+File:button E3.JPG
+</gallery>
-'''1.November'''
-Autoclaved two dishes filled with LA-Medium, one of the dishes filled up with water during the process, discarded and poured with leftover medium that has been in the autoclave, and inoculated with vibrio phosphoreum around 16.00.
+===Artistic Abstract===
-they should start to glow in 12-24 hours.
-'''2.November'''
+To address the growing issue of patented living organisms, I pirate the genome of store-bought mushroom-products, by reverse engineering the product to its viable state of mycelium. I then distribute the organism, for everyone to culture and distribute, in a respectful way and to form mutual diplomatic relations. As the genetic makeup of the strains change through cultivation, the creative commons licence is applied to the genome to keep it from being obscured by companies.
-Pleurotus growth becomes apparent.
-vibrio phosp. still no luminescence. maybe the concentration of the the autoinducer is still to low in the medium. ''(Osamu Shimomura: Bioluminescence, Chemical Principles and Methods chapt. 2.7 p. 42 subchapt. Auto induction)''
-Euglena seem to spin when there is not enough light for them, rather an otherwise. Which makes perfect sense (appetitive behaviour) but somehow I thought it to be the other way round.
+[https://creativecommons.org/licenses/by-sa/4.0/ CC BY-SA 4.0]
-'''8. November'''
+===Scientific Abstract===
-vibrio phosp. dish A shows signs of contamination, I assume due to autoclaving problems as mentioned above.
+Specimen of mushrooms obtained from supermarkets are cloned and the developing strain is purified, than stored in a cold-storage strain-bank. Via reverse-engineering the strain from a commercial product is extracted and cultured. The strains will be given away, and kept available.
-still no luminescence, I am beginning to suspect them to be dark mutants ''(Osamu Shimomura: Bioluminescence, Chemical Principles and Methods chapt. 2.7 p. 43 subchapt. Formation of dark mutants)''
-Pleurotus grow is, if, growing very slowly, planning to move on of the two pegs to another medium.
-[[File:1590301.jpg|400px]]
+===Method===
-supplementary: vibrio phosp. opened jar A and shock it vigorously under the laminar hood, to get oxygen to the bacteria, which should aid glowing. no effect. the sample has to be discarded anyway.
+For every attempt of cloning at least seven petri-dishes filled with PDA- medium are inoculated. The scalpel is sterilized in a hot flame till it glows red. It then is cooled in the receiving petri dish. The mushroom is ripped apart in the middle. Tissue then is taken from just above the gills, inside the stem or just under the cap. It is inserted in the hole that the hot scalpel made. The dishes are labeled and incubated in the incubator (23°-25° C). The mycelium that emerges is purified by running it through subsequent petri dishes till there is no visible evidence of contaminants. Then it is stored in test-tubes filled with PDA- medium at temperatures below 5°C. Two tubes per strain, enveloped in a zip-loc-bag. ( liquid suspended-animation technique will be introduced.)
-'''9. November'''
+===References===
-Discarded jar A of vibrio phosp. due to high contamination.
+*Paul Stamets and J.S. Chilton: The Mushroom Cultivator; Sterile Technique and Agar Culture: Starting a Culture From Live Tissue (p. 29) 1983 Agarikon Press Olympia Washington ISBN: 978-0-9610798-0-2
-transferred on of the pleurotus pegs on malt medium. (Paul Stamets, The Mushroom Cultivator)
+*Paul Stamets: Growing Gourmet and Medicinal Mushrooms (Third Edition); Culturing Mushroom Mycelium on Agar Media: Starting a Mushroom Strain by Cloning (p. 88); 2000 Ten Speed Press ( Random House New York) ISBN: 978-1-58008-175-7
-'''15. November'''
-Pleurotus in malt-medium spawns cottony mycelium. The sample shows mild contamination.
-the tupperware sample still is, if growing very slowly but shows no signs of contamination.
-the temperature of the incubator is now 25.5
-'''16.November'''
-Cooked and sterilised 1 Liter of Potato-Dextrose-Agar-Medium. Poured 5 petri-dishes, to be sterilised on friday.
-Still no luminescence in vibr. phosp.
-'''17. November'''
-sterilised and inoculated 3 petri dishes with Pleurotus strains from the peg on malt-medium. Transferred the original peg to one petri dish and kept one blank for controlling. All dishes are Potato-Dextrose-Agar-Medium.
-the tupperware pleurotus caught on in growing, but still is very restrained.
-'''19. November'''
-Pleurotus: all samples show good growth and no contamination. Cottony mycelium (typical for pleurotus ostreatus) is developing.
-[[File:1590296.jpg|400px]]
-The peg jar looks strange though. I rolled the peg around in the dish on the 17th. Now the surface of the medium shows the tracks of this action.  But the developing organism looks rather yeasty than mycely.
-'''
-.November'''
-All Pleurotus samples look very promising. The F1 generation samples are thriving. No contamination at all. Very soon the production of spawn can begin. The peg sample is now all cottony mycelium. So no yeasts, just the look of them. Maybe there are more of this early stage structures in the sample, or it is minor contamination. The control blank shows no sign of contamination.
-[[File:1590297.jpg|400px]]  [[File:1590300.jpg|400px]]
-Inoculated on jar of LA-bacteria-medium with kirby and cabbage kimchi samples.
-'''29. November'''
-The kimchi jar shows no signs of growth, nor contamination. Nothing shows up under the microscope.
-'''30. November'''
-Prepared rye for grain master according to Stamets.
-'''1. December'''
-Inoculated Grain Master G1. (2x 150g)
-'''3. December'''
-Placed one of the G1 Masters in the incubator at 25 C.
-'''5.December'''
-G1 Masters are running.
-Autoclaving dark mutants and empty kimchi jar.
-Prepared  six G2 bottles according to Stamets (soaking overnight to germinate endospores)
-'''6. December'''
-sterilised 3 of the G2 and inoculated with G1A.
-[[File:stirrer.MOV]]
-I built an magnetic stirrer. It runs on 5 volts. Should be sufficient for now.
-'''7. December'''
-sterilised the other 3G2
-Two Pleurotus samples were dried out in their dishes in an attempt to let them form primordia, with they did, until they where hard and dead.
-They might be of value to Suna, and her paper project, I keep them.
-'''8. December'''
-Inoculated 3 G2B.
-'''9. December'''
-Grain 2 Masters running! Marked G2B.
-G2A mycelium is getting a pinkish tone!
-Inoculated 6 Potato Dextrose Agar dishes with Brown Button Mushroom
-Method: cloning. Strain: commercial, Poland ( from the supermarket)
-'''13. December'''
-Button Mushroom cloning. dishes showing mycelial leap of on the medium: A. B. C. D. E.
-Dishes showing no mycelial growth whatsoever: G.
-shock all G2 Masters.
-One of the G2A jarls lost its professional filter cotton ball. I employed questionable practices and replaced the cotton ball after extraction under the laminar flow hood, with a new one I soaked in ethanol.
-I separated the jar from the others and marked it with a red circle.