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Topics - Polybius

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PF - Tek / None of my spore prints worked
« on: October 23, 2013, 03:28:42 PM »
I inoculated about 40 jars using syringes I made from prints from my first grow and none of them have colonized. One of them has four quarter sized white patches of mycelium which appeared overnight, but that one has completely ceased it's growth. Not a single one of any of the others shows any signs at all of growth, and it's been three weeks now.

I prepared the substrate the exact same way I did last time. I'm very certain the substrate isn't the issue, because I did a test with a jar that I did not sterilize, and it's beginning to grow some sort of random white mold inside it. So clearly the substrate allows for some level of growth.

I thought that maybe the water levels of the substrate were off, or something, but out of forty jars, you'd think that that would mean that some would simply colonize faster than others, or not at all.

I also experimented with using a small amount of spore solution compared to a lot of spore solution. On two jars, I used an entire syringe. Nothing.

This led me to believe that the problem had to be with the spores themselves. I researched how to make a liquid culture to test further. Sterilize a jar, boil water, fill the jar with the water, mix in about a tablespoon of clear honey, and then add a few cc's of spore solution. Every report I've read online indicates that spores in this environment will produce mycelium with a matter of days. Guess what. Nothing.

So I've narrowed the issue, I THINK, down to the spores themselves. Now, clearly SOME of them are still viable. One single jar had growth before it stopped. The thing is, I used that syringe on many other jars, and they produced nothing.

I made spore prints on both card stock as well as tin foil, and have used both those to make syringes, so I don't think it has to do with what I made the print on. The only thing that I failed to do when making the prints was letting them dry for two days after removing the caps before wrapping them in foil. COuld this be the issue?

I can only assume that the issue is something to do with the spores. When I made the syringes, I made sure to boil the water for at least ten minutes to get rid of impurities. And I let the syringes filled with water cool in the fridge to make sure that hot water would not damage the spores. But, as I have said, nothing.

I made spore syringes two ways: the first way was to boil the glass that I used to spray the water from the syringe into. The second way was your suggestion: to wipe it with alcohol. I did this all inside a glove box.

Most of my syringes I let hydrate for 24 hours, but the first one I did not. No difference.

Here are my working theories:

1.) Maybe there was still alcohol floating in the air inside the glove box that damaged the spores when I transferred them from the prints to the glass.

2.) Maybe the boiled tap water has chemicals in it which kill the spores when they are put into the syringe. (However, if that were true, shouldn't the water I used to mix the substrate also kill them when they are inoculated? Maybe not, because the jars are sterilized for an hour and a half, which may breakdown whatever chemical is preventing their growth. I don't now.)

3.) Maybe all of my prints were rendered nonviable somehow during the printing process. But if that were the case, not a single one should have produced growth, yet one did.

4.) God hates me.

The best I can come up with is to try making a liquid culture in a very unsanitary way, by simply scraping the spores directly off of a print into a jar of honey water. Maybe that will let me know if the issue has something to do with the syringes or not. The only other thing I can think of is to buy a bottle of distilled water, make syringes with that, and see if it makes any difference. I'll be experimenting with both.

Any help here would be greatly appreciated.

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