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spore print to agar


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#1 LIFELESSDEAD

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Posted 09 September 2020 - 02:33 AM

yay, so I just got my GT spore print. Ive read alot about spore print to syringe then to whatever but not much is said about spore prints directly to agar. Can I go directly to agar? what would be the problem with doing so? too many spores competing?



#2 ElrikEriksson

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Posted 09 September 2020 - 03:22 AM

If the print is clean and your agar technique is good there should be no problem. Spores are the normal way to start a culture on agar.

If the print is dirty or your technique is poor you'll have lots of work transferring away from contams.

HERE you can see me spreading the spores thinly enough to isolate monokaryons without magnification, so too much competition is only an artifact of spreading technique. You can spread them thinly, and even breed your own hybrids if you own a microscope that gives a clear view at 400X.


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#3 LIFELESSDEAD

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Posted 09 September 2020 - 04:34 AM

cool, thanks. awesome info. My plan is to take a inoc loop then and inoculate to agar then. So should I still go for, less is more, even though i want dichariotic?



#4 ElrikEriksson

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Posted 09 September 2020 - 11:43 AM

That's what I usually do, though I have been surprised by prints of especially low viability.

Sometimes what I do is to streak 2 plates very thinly and one plate with more just in case. Then when I'm ready to go to grain I don't do wedges, I put 1,5 to 5,5 ml of sterile distilled water on a contam free plate and scrub the mycelium into the water before sucking up into a sterile syringe and injecting to jars, 1 ml per pint jar. Obviously I'm not a big producer :laugh: I just grow out 2 dubtubs of each variety so I can try out more varieties and hybrids, because for me when dosed at 7 grams different varieties do feel different.

 

There have been times I scrubbed one side of a plate into water for one syringe and the other side for a second syringe. My technique must be adequate because they didn't contam.


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#5 LIFELESSDEAD

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Posted 09 September 2020 - 08:36 PM

So I kinda screwed up but I just scraped a few spores with m yh scalpel and dumped them on agar. I feel like they clumped a little. Hopefully all will be ok. Perhaps I should go into the dish a try to spread them or just let them be and try again?

#6 ElrikEriksson

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Posted 09 September 2020 - 11:34 PM

I'd let that one be and you can also start another if you like.

When you're new to agar every opening of the dish runs a high risk of contam.

 

A great way to get better at agar is to pay attention to everything every time you do any work while trying to move as little as possible.


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#7 rockyfungus

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Posted 10 September 2020 - 06:07 PM

I just scrapped a ton of spores to a ton of different agar.

Prints are not inherently clean so best to get some spores onto the agar and start transferring to new plates as you can identify clean growth.

I just put a clumpy print to agar and it didn't mater, tons of fuzz to choose from and 1 contaminate to avoid.

Spores will take 4-5 days usually to germinate on agar as well.


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#8 peacefrog

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Posted 11 September 2020 - 07:49 PM

As stated above, yes spores to agar will work perfectly. If you had a clump of spores on the surface of the media, no problem. Remember spores are microscopic, so even if you don’t see any signs of spores on your loop, there are probably hundreds or thousands of them on there, so you don’t need clumps. But if it happens, it happens. Just let it ride and see what happens.

I’ve done countless agar plates/jars and have seen that happen countless times. Sometimes I even encourage it with old spores. Just wait it out to see what germinates, mycelium or contamination. Like said above, streak more plates. That will only exponentially add to your success. A lot of this hobby boils down to law of averages IMO.

And spores on agar will germinate 2-14 days later, more or never. I have seen them germinate within 2 days and I have seen them do the same in 10-14 days, or nothing. It just depends on several factors. Age of the spores, cleanliness, temperature, media, proper technique etc.

But if there is only a tiny point of clean mycelium on the plate, you an easily clean it up by more transfers to agar.

Just my 2 cents.
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#9 rockyfungus

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Posted 12 September 2020 - 05:25 PM

Great advice above.

I like to try and just get enough spores to land in maybe 2-3 places. Sometimes your agar is just black spotted, don't really matter.

My biggest thing I need to work on is saving those spore prints! I tend to try and use every last spore and make like 20 plates off of the print. Which will snowball into 60 easily once you take 2-3 swipes from each plate you printed.

On a plate with spores it can be very hard to identify what the hell you have. Just try and grab the teeniest speck of some clean white fluff and after a transfer or 2 you should know if you have mycellium or contams.

Usually by T1 I can go to grain, but prefer to go out to T2 to make sure I didn't miss any contams.



#10 LIFELESSDEAD

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Posted 12 September 2020 - 06:33 PM

more great advice. So, I put some dust from a baggy on to agar and i apologized i did not document, but thanks to advice and waiting like 2 weeks and i got growth, so yes your right it doesn't take much to grow something. Im surprised i didnt get more contamination. I have some red dots and the rest looks like growth maybe a few fuzzballs but they dont seem to colonize very fast. pic coming before i throw it out!



#11 LIFELESSDEAD

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Posted 12 September 2020 - 07:43 PM

Still slow growing mycelium mixed with mold right?

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#12 Uncle G

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Posted 12 September 2020 - 08:19 PM

Very interesting.     



#13 peacefrog

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Posted 13 September 2020 - 01:40 AM

Still slow growing mycelium mixed with mold right?


Sure looks like it. I see you have transferred a few times from that plate. How are those plates looking?

#14 rockyfungus

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Posted 14 September 2020 - 04:29 PM

Never seen that flesh colored colony before. Kinda odd that it looks like myc with it.



#15 LIFELESSDEAD

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Posted 15 September 2020 - 11:50 PM

Still slow growing mycelium mixed with mold right?

Sure looks like it. I see you have transferred a few times from that plate. How are those plates looking?

Here's one for sure, unfortunately I'm realizing my documenting technique sucks I'm starting to lose track of which plates came from what. No big deal for this first round as long as i learn, Hopefully i don't mix up my store cloned crim with my spore germed cube lol

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#16 Arathu

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Posted 16 September 2020 - 04:57 AM

 

 

Still slow growing mycelium mixed with mold right?

Sure looks like it. I see you have transferred a few times from that plate. How are those plates looking?

Here's one for sure, unfortunately I'm realizing my documenting technique sucks I'm starting to lose track of which plates came from what. No big deal for this first round as long as i learn, Hopefully i don't mix up my store cloned crim with my spore germed cube lol

 

You would not be the first nor will you be the last to see surprise fruiting from such things......get Sharpies or grease pencils (both in fact)

 

Looking good so far man......keep working them....

 

A






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