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Offlinejords
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Registered: 02/23/20
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first try in about 15 years... agar and other questions
    #26682516 - 05/20/20 03:49 AM (3 years, 8 months ago)

well, it's my first try since i did a bunch of PF cakes a really long time ago.
i'm really trying to do it right this time so i got myself read up and outfitted for agar.

i'd bought some syringes more than a year ago (when i first felt the inspiration to go for it again). PE, B+, golden teacher, treasure coast.

i did the inoculations in an sab. i did 4-6 plates for each variety, most i just put a drop in the middle, but i tried a streak plate for each one - not a zig-zag, but the classic kind where you kind of extract into different sectors to try to isolate things. i wrapped em in saran wrap and kept the temp in the low 70's.

so... it took about 10 days to see some growth -- luckily almost all mycelium

i'm at day 20 now and the results are like this:
3/5 TC
1/6 PE
3/4 B+, but the streak plate has sporulating mold.
0 GT germination. streak plate has sporulating mold on it.
aside from the mold i see tiny little contam in only one germinated plate and 1 or 2 un-germinated.

i'll post some photos below if i can figure that out. but here's my laundry list of questions that have come up in the process so far. any feedback would be so helpful. i know most of these answers are somewhere on this site, but fuck, i've done so much reading i'm kinda burned out on reading threads and also i thought maybe it's time to make a post and enter the community.

so is it normal to take 10 days to germinate? what's average?
why would i have so many blank plates (like 12 of them)? do spores lose viability in a year? they had clumped up in the syringes, but i shook the shit out of those things before using them. so not sure if its spore viability, or there were just no spores in the drop. also i cooled the syringe tip by squirting some out before dropping, but is there any possibility of the tip still being too hot and killing spores? hardly matters, i'm hoping to never buy or use a spore syringe again. but still curious. i do want to get some of each variety, so i may squirt some more PE and GT -- any reason not to re-use the clean ungerminated plates?

i've been looking at the plates just about every day, and restacking them willy nilly and the next day the ones on top always have condensation. anyway to just not have condensation? is it better to stack upside down?

for my first agar work - inoculating 4 varieties, i got a little confused about order of operations as far as wrapping and labeling and keeping everything in order. any advice? i mean, it all worked out, i'm just looking for those elegant solutions from pros. do you wrap outside the sab or in? how about labelling? what are the key stats to put on the plate, and how do you keep it organized? do you keep track of transfers? do you keep track of the whole ancestry? or is that obsessive?

once i make the first transfer what do i do with plate that i just cut all up? get rid or save?

when you're slicing a plate into wedges for a grain transfer, let's say 8 wedges, would you sterilize your blade before each of the 4 cuts? or just once at the beginning? or cut each wedge individually?

is there an ideal time to do agar to agar transfers as far as the myc is concerned? ie how big should the parent specimen be? when taking transfers from the leading edge, is the leading edge the mycelium edge itself? or does it refer to the mycelial "halo" that extends about 1/8" in front of the white mycelium?

does anybody use agar w/ hydrogen peroxide?

why do people bother with microwaves for making agar? what's the benefit over just cooking it in a saucepan on the stove?

only one PE plate germinated and there are just 2 sad little 1/2" dots of mycelium on it after 20 days. what's up with that?

...
ok that's a few questions. i thank anyone who reads through all of this. this site is amazing. 20 years ago i was in college and it helped me to grow some mushrooms in my closet. it's really incredible all of the progress in techniques that's been made since then, and the breadth of knowledge that's recorded here. thanks everyone for their efforts.


this one has a tiny contam just below the T and C


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OfflineSockadin
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Registered: 01/03/10
Posts: 7,244
Last seen: 2 months, 20 days
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Re: first try in about 15 years... agar and other questions [Re: jords]
    #26682686 - 05/20/20 06:48 AM (3 years, 8 months ago)

Damn that's a list.

I wouldn't reuse plates.
I don't flip my unused plates.
Take the smallest transfer possible. Look up Demonic's hole punch Tek.
I don't cut wedges, once I have gone T4-5 I will use the whole.culture if it is 50-60% colonized to knock up jars.

Yes spores do loose viability over time but it can also take 30 days or more with spores that old.

Good luck, hope I got all the questions.


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