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goldcap
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reviving +15 year old spores
#27864522 - 07/16/22 08:14 AM (2 years, 5 months ago) |
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Hi.
As the title has it, this is a documentation of reviving +15 year old spores. The spores were stored sandwiched between two pieces of glass, fastened by tape. Stored in darkness in room temperature.
(Started off with like 30 prints, like 3-4 of them germinated in honey LC but mold killed off the mycelium. Now an attempt with the last good print...)
Spores were introduced to LC in a home made sterile glove box. Zero contamination so far. The jars with spores have been in 26-28 degrees, darkness for a few weeks so far. No sign of germination yet.
The liquid culture used was: 100 grams of potatoes boiled in 1 liter water until soft, filtered until liquid was more clear. 2 (European) table spoons of dextrose were mixed into the liquid and then sterilized in pressure cooker for 30 minutes.
Besides these pure liquid LC jars, some of the LC was mixed with Agar (1.5 table spoons of agar per 1 liter water) and cooked for 2 minutes before sterilization. Spores scraped with sterilized knifed and "cut" into the agar. No germination on the agar either.
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Land Trout
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Re: reviving +15 year old spores [Re: goldcap] 1
#27864539 - 07/16/22 08:45 AM (2 years, 5 months ago) |
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Edited by Land Trout (07/16/22 09:25 AM)
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goldcap
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Re: reviving +15 year old spores [Re: Land Trout]
#27867134 - 07/18/22 06:29 AM (2 years, 5 months ago) |
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thanks. this give me great hope
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DERRAYLD
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Re: reviving +15 year old spores [Re: goldcap]
#27867210 - 07/18/22 08:00 AM (2 years, 5 months ago) |
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I've recently germinated Puerto Rican from 04 that I've been isolating further, I used a softer agar and soaked swabs I made from spore prints in sterilized water.
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goldcap
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Re: reviving +15 year old spores [Re: DERRAYLD]
#27954382 - 09/17/22 10:56 AM (2 years, 3 months ago) |
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Ok, good tip!
I ran out of spore prints, but in a desperate attempt I took some of the liquid culture and pored it over the agar. About 1-2mm of culture now covering the agar surface.
No success. No growth, not even from contaminants ;-).
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bakedbeings
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Re: reviving +15 year old spores [Re: goldcap] 2
#27954394 - 09/17/22 11:02 AM (2 years, 3 months ago) |
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spores to LC seems like not the best way to do this. you are eliminating your ability to separate clean growth and instead creating an all or nothing scenario, which in the case of spore LC is almost always nothing
-------------------- Confused? Well now you can!
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goldcap
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Re: reviving +15 year old spores [Re: bakedbeings]
#27999886 - 10/15/22 04:59 AM (2 years, 3 months ago) |
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Hoooray, there is a cloud... but first:
Quote:
Bakedbeings said: spores to LC seems like not the best way to do this....
Yes I agree. I have now studied and made some agar. That did not work either. So I tried with "forced hydration" using a syringe and rubberbands... but well, no success so far.
However, when I was making room for the agar I removed a LC jar and put in my closet (so I could procrastinate cleaning a little longer) and what do you know, today there is a cloud in the LC. :-O
While the closet is dark most of time, every once in a while there door is open and/or the light is turned on.
The temperature is like 21 Celsius.
So the same jar that stood in min 25 to max 28 Celsius (most of the time 26 Celsius) and nothing happened, when I put it in 21 Celsius suddenly things started to happen.
Maybe a coincidence? Maybe not.... the spore print is 15-16 years old, so I will have to wait like 15-16 years to repeat this experiment.
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goldcap
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Re: reviving +15 year old spores [Re: goldcap]
#27999890 - 10/15/22 05:04 AM (2 years, 3 months ago) |
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Ok... so now I have a cloud in the LC.
I should probably remove the cloud from LC to agar right?
What is the best way to go about it?
I am thinking that maybe I could like sterilize a spoon, scoop up the cloud and throw the thing onto an agar plate! Sounds good?
Another alternative could be to try and suck it up using a syringe...
Any other ideas?
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bakedbeings
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Re: reviving +15 year old spores [Re: goldcap]
#28000021 - 10/15/22 08:07 AM (2 years, 3 months ago) |
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yes, get the myc onto agar, however you do it
i would go with water agar. or alternatively a brf puck
-------------------- Confused? Well now you can!
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goldcap
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Re: reviving +15 year old spores [Re: bakedbeings]
#28000044 - 10/15/22 08:42 AM (2 years, 3 months ago) |
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Thanks for the advice!
Quote:
Bakedbeings said: yes, get the myc onto agar, however you do it
i would go with water agar. or alternatively a brf puck
I was not aware of water agar. After doing 1 minute of research, I have learned that the basic idea is to starve bacteria of nutrients (water agar is made from distilled water) while the mycelium retain some of the nutrients from the original LC (plus some of the LC on the surface of the mycelium).
This way the healthy myc can be isolated from any bacteria.
Hmm... I like only got ONE shot at this, because I have no other spore print, so I really want to do this right.
The cloud is maybe 1-1.5 cm in diameter (perfectly circular). I will give it some time to grow out and become bigger, before I take a part of it and move to agar.
I will take one part to water agar. And another part to nutrient agar. And then keep rest in the LC.
Edited by goldcap (10/15/22 08:43 AM)
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Rusty2096
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Re: reviving +15 year old spores [Re: goldcap]
#28000049 - 10/15/22 08:45 AM (2 years, 3 months ago) |
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You have many many many shots at this - not only 1 like you said. Suck up that cloud in a syringe. Squirt 1 drop per plate (you can do both some WA plates and nutes plates)
-------------------- Currently looking for nothing. You guys who sent me stuff are straight up awesome!.
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Semi-solid liquid culture (SSLC)
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Vibetyme
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Re: reviving +15 year old spores [Re: Rusty2096] 2
#28000150 - 10/15/22 10:13 AM (2 years, 3 months ago) |
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When you have 50 posts check out the marketplace for free spores. It will show up toward the bottom of the thread list.
People around here are very very generous with their work.
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bakedbeings
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Re: reviving +15 year old spores [Re: Rusty2096]
#28000154 - 10/15/22 10:16 AM (2 years, 3 months ago) |
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Quote:
Rusty2096 said: You have many many many shots at this - not only 1 like you said. Suck up that cloud in a syringe. Squirt 1 drop per plate (you can do both some WA plates and nutes plates)
-------------------- Confused? Well now you can!
HHG - cheapest way to start - how i roll
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goldcap
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Re: reviving +15 year old spores [Re: Rusty2096]
#28001416 - 10/16/22 06:18 AM (2 years, 2 months ago) |
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Quote:
Rusty2096 said: You have many many many shots at this - not only 1 like you said. Suck up that cloud in a syringe. Squirt 1 drop per plate (you can do both some WA plates and nutes plates)
Ok... that sounds really really good!
I was not aware that 1 drop is enough per plate, so thanks for telling me!
Also thanks for your advice on using a syringe! I will do this create both water agar and agar with nutrients.
Pretty wild if its possible to bring back 18<= this many year old >= 15, spores. They were stored in room temperature, darkness and printed on tinfoil, which were stored in a ziplock bag.
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goldcap
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Re: reviving +15 year old spores [Re: bakedbeings]
#28001418 - 10/16/22 06:23 AM (2 years, 2 months ago) |
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Quote:
Bakedbeings said:

Cool! I will follow ur advice!
I am curious as to why the spores germinated when I put them in 21 Celsius degrees, while nothing happened in 26.
(Hmm, I mean it could be that it is not cubensis myc growing here)
But to be perfectly accurate the jars was _first_ placed in 26 celsius and after +2 weeks moved to 21 Celsius, so maaaybe it germinated under warmer conditions and grew out in the colder.
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goldcap
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Re: reviving +15 year old spores [Re: goldcap]
#28001467 - 10/16/22 07:45 AM (2 years, 2 months ago) |
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Ok... I followed an old tek, so I read a little about temperatures and now I realize that the temperature I was aiming for was like far to high.
21 Celsius seems to much close to ideal growing conditions.
Apparently 23 seems to be very good temperature to have, because you should be able to have that through the whole grow process. While 24 seems to be the best, the actual temperature in a colonized jar will be slightly higher because the myc produce heat... and the less warmth, the less contaminants... 23 seems to be a sweet spot if you dont ever want to change temperature.
Also I was thinking about how to optimize the germination of super old spores using a ghetto setup. I am thinking that forced hydration using pure water... wait like 2 weeks and then make a LC and put a few drops of the spore water in the LC.
Since you can create an almost endless amount of LC jars from the sporewater, there is plenty of jars that can develop myc. If there are contaminations, the myc can be easily moved to agar for isolation.
The problem I have had with applying spore water directly onto agar, is that the spores looks too dry. In LC they can all the water it can possibly need... sometimes it takes like several weeks for this old spores to germinate. I had one print that was 15 years old, put it in a LC and after 3 months there was germination (that particular grow failed during colonization because of bacteria or something).... I mean if you like rrreally need old spores to germinate (and have time) you can do this. The agar surface seems to dry out after several weeks.
Also you save some agar... when you do the LC you always end up with more LC then you need anyway, so with many jars you can make use of all the LC.
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goldcap
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Re: reviving +15 year old spores [Re: goldcap]
#28001470 - 10/16/22 07:48 AM (2 years, 2 months ago) |
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I wonder how many newbie-no-nothin-wanna-be-growers (like me) find themselves having the problem of reviving decades old spores... can not possibly be that many.
Edited by goldcap (10/16/22 07:49 AM)
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Rusty2096
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Re: reviving +15 year old spores [Re: goldcap]
#28001490 - 10/16/22 08:12 AM (2 years, 2 months ago) |
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Quote:
goldcap said: I wonder how many newbie-no-nothin-wanna-be-growers (like me) find themselves having the problem of reviving decades old spores... can not possibly be that many.
1 - you
The other noobs buy grow kits 
Quote:
21 Celsius seems to much close to ideal growing conditions
21 is perfect. Higher temps you were maintening (like 26) is still inside cubes myc acceptable range but the thing is, at that temp, you will favorise bacterial growth.
Spores can take up to many weeks to germinate (especially older ones). While moving to 21c was the right thing to do , I doubt that was the turning point for germination. More likely a coincidence.
-------------------- Currently looking for nothing. You guys who sent me stuff are straight up awesome!.
We don't own things - things own us.
Semi-solid liquid culture (SSLC)
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SymPlayTon
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Re: reviving +15 year old spores [Re: Vibetyme] 1
#28001515 - 10/16/22 08:32 AM (2 years, 2 months ago) |
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Quote:
Vibetyme said: When you have 50 posts check out the marketplace for free spores. It will show up toward the bottom of the thread list.
People around here are very very generous with their work.
I am still waiting for marketplace access lol.
Still in the 90 day waiting period lol
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SymPlayTon
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Re: reviving +15 year old spores [Re: goldcap]
#28001518 - 10/16/22 08:36 AM (2 years, 2 months ago) |
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Quote:
goldcap said: I wonder how many newbie-no-nothin-wanna-be-growers (like me) find themselves having the problem of reviving decades old spores... can not possibly be that many.
I would guess not too many lol.
I would assume the process is the same. The big difference being more patience and more finger crossing.
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bakedbeings
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Re: reviving +15 year old spores [Re: SymPlayTon]
#28001525 - 10/16/22 08:46 AM (2 years, 2 months ago) |
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OP is playing mush cult on hard mode
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SymPlayTon
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Re: reviving +15 year old spores [Re: bakedbeings]
#28001535 - 10/16/22 08:57 AM (2 years, 2 months ago) |
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He is just using it as a flex so other newbs like me can't complain.
Edited by SymPlayTon (10/16/22 08:57 AM)
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goldcap
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Re: reviving +15 year old spores [Re: Rusty2096]
#28004547 - 10/18/22 11:24 AM (2 years, 2 months ago) |
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Quote:
Rusty2096 said:
Quote:
goldcap said: I wonder how many newbie-no-nothin-wanna-be-growers (like me) find themselves having the problem of reviving decades old spores... can not possibly be that many.
1 - you
The other noobs buy grow kits 
Quote:
21 Celsius seems to much close to ideal growing conditions
21 is perfect. Higher temps you were maintening (like 26) is still inside cubes myc acceptable range but the thing is, at that temp, you will favorise bacterial growth.
Spores can take up to many weeks to germinate (especially older ones). While moving to 21c was the right thing to do , I doubt that was the turning point for germination. More likely a coincidence.
I dont believe in coincidences, so the most likely reason is that on the same day I noticed growth,I also shaved my legs and drop my computer on the ground... yeaaa. that have to be it!
But hey, yea probably I was just damn lucky. Good point about keeping temperature lower to reduce contaminates! I will keep it lower!
I did lie a little bit though... I did grow some shrooms like 15 years ago so its not the first time ever. I ate them maybe 2 times and made 30 prints or something and then never thought of it again.
Out of these 30 prints, I have gotten 2 previous ones to develop into clouds. But since I am well basically a n00b for all intents and purposes, I used the most horrible growing techniques and contaminants had my cubes for breakfast.
Now I got no more prints but one jar with a cloud, lol. Lucky bastard right! 
My plan is to suck up the myc cloud from the LC and drop it onto several agar plates.
Then if the myc grows I will cut out healthy agar and move onto other agar plates, so they I get plenty of plates.
Then I guess I will wait for one of the agars to be fully overgrown by myc, before attempting to inoculate a jar with grains.
I am aiming for getting from myc to mushroom ASAP... so I get can some fresh spores....
...then I will wait 30 years and see if I am able to germinate those 
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goldcap
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Re: reviving +15 year old spores [Re: SymPlayTon]
#28004563 - 10/18/22 11:34 AM (2 years, 2 months ago) |
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Quote:
Bakedbeings said: OP is playing mush cult on hard mode
Quote:
SymPlayTon said: He is just using it as a flex so other newbs like me can't complain.

Well I lied a little. I did make the spores myself all those years back. I did a handful of grows and made some spores. Not super n00b. But clearly I am at least making n00b mistakes here, so well... 
I made spore prints sandwiched between glass and taped up so no air could get inside. Stored in ziplock. Darkness. Room temperature. Perhaps 2 of those prints produces clouds in LC.
This last print was made on tinfoil, the edges of the print folded so no air could get inside. Stored in ziplock bag. Room temperature.
Apparently tinfoil worked basically as well as glass, for preservation. But they can easily get damaged by accident, tearing a whole.
N00b or not, I hope this thread will be helpful for other people.
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SymPlayTon
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Re: reviving +15 year old spores [Re: goldcap]
#28005649 - 10/18/22 11:24 PM (2 years, 2 months ago) |
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I am excited to keep up with your results and see your grows!
I hope to learn a lot from your logs.
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goldcap
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Re: reviving +15 year old spores [Re: SymPlayTon] 1
#28006178 - 10/19/22 10:06 AM (2 years, 2 months ago) |
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Quote:
SymPlayTon said: I am excited to keep up with your results and see your grows!
I hope to learn a lot from your logs.
Smart move! If you follow me closely I can make all the mistakes so you dont have to 
I will keep the thread updated of my results.
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goldcap
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Re: reviving +15 year old spores [Re: goldcap]
#28006241 - 10/19/22 10:46 AM (2 years, 2 months ago) |
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How to sterilize syringe?
I tried boiling it (PC), but it damaged the syringe (the plastic went bad and water leaked out when pressing the plunger).
I dont have a UV lamp strong enough to kill bacteria...
But I am thinking that maybe I could suck up some chlorine, leave it in the syringe a little while and then empty the syringe. If I wait like 7 days the chlorine drops still remaining should have evaporated?!
Are there any better tricks?
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bakedbeings
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Re: reviving +15 year old spores [Re: goldcap]
#28006256 - 10/19/22 10:51 AM (2 years, 2 months ago) |
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not sure what went wrong but i sterilize syringes in the PC all the time. i reuse the same ones for years and they still work. they are the originals from when i bought spore syringes
ive heard of people simply boiling water and pulling it into the syringe, pushing it out, and repeating several times. gets the thing pretty clean i guess. but i dont see why you cant do it in the PC
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goldcap
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Re: reviving +15 year old spores [Re: bakedbeings]
#28007309 - 10/20/22 01:18 AM (2 years, 2 months ago) |
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Quote:
bakedbeings said: not sure what went wrong but i sterilize syringes in the PC all the time. i reuse the same ones for years and they still work. they are the originals from when i bought spore syringes
ive heard of people simply boiling water and pulling it into the syringe, pushing it out, and repeating several times. gets the thing pretty clean i guess. but i dont see why you cant do it in the PC
Thanks for your input and this confirms what I have read elsewhere.
My first idea was to PC the syringe and I tried it, then tested it and noticed that it stopped working.
My bet is that your syringe has an O-ring at the bottom of the plunger, that is slightly bigger then the inside of the syringe, so that the O-ring is pressed against the sides. That way no water can leak out when you apply pressure on the plunger.
My syringe has no O-ring. Instead it is relying on having like a perfect fit to the inside of the syringe. I noticed this at once when I got the syringe and was confounded "will this really work?".
Actually it works really well. I can even put rubberbands (forced hydration) on the plunger, which will slightly bend the top of the plunger (where you put would out finger when pressing) but no air or water is able to leak out, between the plunger and the inside how the syringe.
After having PC the syringe, its no longer air or water tight. My bet is that the high temperature deformed the plastic (I dont believe in magic and dont think nano technology played any part in this particular case.)... has got the be the heat.
So therefor I got 2 ways to go with sterilizing without heat: UV-light chemicals
UV-light is nasty stuff... can easily damage your eyes and stuff. I dont want that crap lying around in my tent, should a cat or some of the nature people wander around and find it and look into it by accident.
Chemicals: I got isopropanol and chlorine. Both are easily obtainable. Isopropanol can be found in most disinfectant. Even hand disinfectants could maaaaybe work. They usually have some glycerine, but that actually only a benefit because it will make it less likely for the spores to attack to the syringe itself. But there is other crap in it as well, such as some kind of thickeners chemicals and I dont know how that will affect the spores. Then there are car cleaning stuff that contains isopropanol, but who knows what other kinds of chemical residues might be there... I mean its not food graded ;-)
However, I already got some chlorine at hand and hey, accordingly to internet it breaks down in a few days (not as fast as isopropanol though). Especially if the sun (UV) is shining on it. Less of a fire hazard too...
So rinsing the syringe with water, then sucking up some chlorine and then putting it in the sun to dry out... that should work. Chlorine is not food graded either, there will be some toxic stuff left from that too.
Hmm... maybe rinsing out the syringe with chlorine then cleaning it out with pre-boilied water would work. That way there would be no chemicals left. Yea thats probably it.
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goldcap
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Re: reviving +15 year old spores [Re: goldcap]
#28025559 - 10/31/22 03:00 AM (2 years, 2 months ago) |
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Update: All the agar plates have a really healthy and thriving green mold looking ugly stuff growing on them. It looks like the cloud I thought was cubensis, was anything but. That was my last spore print, so end of this test. Fun as long as it lasted.
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