natureboygrower
Well-Known Member
I think taking a couple days off from microdosing is a good idea. Might help reset things a bit so tolerance doesn't increase too dramatically.
Anybody growing mushrooms willing to talk???
- Thread starter 7L!fTeD24
- Start date
Aeroknow
Well-Known Member
Yeah. I don’t think you’re supposed to do it continuously.
I did it for a month, my buddy makes them from my strains, i did feel refreshed but i also take enough to trip as often as i can.
But i’d treat micro dosing like shrooming. You take a gram one day. It’s gonna take 2 grams the next day. So maybe 30 days on 30 days off? Then 30 on?
Or just trip balls once a week. That’s about what i do lol.
canndo
Well-Known Member
Yeah, suppress inoculations are pretty hit or miss
Aeroknow
Well-Known Member
What is your go to method when you are dealing with spores just curious.
canndo
Well-Known Member
Agar dishes and then sector selection then another dish then liquid culture then test.
Aeroknow
Well-Known Member
Nice.
Question. From a swab/spore print, that’s what i do. From a spore syringe you do the same?
Just seems like in a 10cc spore syringe there really isn’t much spores to squirt out on an agar plate. One drop?
canndo
Well-Known Member
Yes, one drop is plenty
Drop That Sound
Well-Known Member
You could also squirt some of the solution out into a clean shot glass, and use an inoculation loop to stir around in it, and then streak it across the agar surface in a small Z pattern. Or use the T or quadrant methods, etc, which further Isolates any possible contams by spreading it around into multiple different sectors, getting more and more thinned out for each one as you go.
canndo
Well-Known Member
You can't do sector selection if you swab the dish. Spores need to find complementary spores and make clamp connections. The closer the spores are together the quicker they select their mates and begin mycelium migration across the plate. So you place your drop dead center of the dish.You could also squirt some of the solution out into a clean shot glass, and use an inoculation loop to stir around in it, and then streak it across the agar surface in a small Z pattern. Or use the T or quadrant methods, etc, which further Isolates any possible contams by spreading it around into multiple different sectors, getting more and more thinned out for each one as you go.
One can often find distinct sectors from that first inoculation and then ensure distinctions with a second transfer from somewhere near the edge of the plate. This means the mycelium has had a few inches in which to organize and differentiate.
This swab technique is not for such a process. If one is attempting specific genetic transfer, the spore solution should be carefully diluted by halves until you suspect that there are only a few single spores in a ml of water. Then use this to spread the few spores as far apart as possible Using that whole dish swab.
Now you can work with the few colonies that emerge.
Drop That Sound
Well-Known Member
I thought the purpose of streaking the patterns with a loop was to spread as many possible contaminants away from the spores over the surface as possible. The thin loop picks up just the right amount of spores you need, without the extra water if you just dip it in. You can also smear out a drop the same way too.
All MSS should be considered non sterile as is right.. so why not go ahead and streak your first plate every time that way? I suppose it would be the best method if you knew your spore solution or print was in fact contaminated, and trying to salvage anything you can.. In the above gif, you would only transfer from sector 5 towards the middle, which would be the most thinned out. More likely to find something clean to take from in that area right? Then, transfer it to the middle of a new plate as usual...
All MSS should be considered non sterile as is right.. so why not go ahead and streak your first plate every time that way? I suppose it would be the best method if you knew your spore solution or print was in fact contaminated, and trying to salvage anything you can.. In the above gif, you would only transfer from sector 5 towards the middle, which would be the most thinned out. More likely to find something clean to take from in that area right? Then, transfer it to the middle of a new plate as usual...
Drop That Sound
Well-Known Member
canndo
Well-Known Member
canndo
Well-Known Member
Essentially true but it still depends. If I have contamination I "race" the mycelium. This involves germinating a few spores and then placing another "slab" of agar on top sandwitching the grow between the agar. This limits the growth to two dimensions. If the contamination is bacterial (which it most often is) the bacteria will not travel but the mycelium will grow away from the infected spot.
You can then simply cut a small block of pure mycelium out and place it on a new dish. Poof. You have isolated the mycelium from the contam.
If that contam is mold the mold will grow at a different rate, usually slower. You can eventually differentiate the two different mycelium and carefully cut out the one you know to be the good one.
It is usually obvious especially with Cube mycelium.
I use my phone flash as a light source beneath the dish, circle the good spot with a red marker on the bottom of the plate to ensure I get it.
I use this most often though with samples from the wild. Clearly there will be contams in there after the sterilized loop is plunged into the gills.
I rarely get contam from syringes.
Chunky Stool
Well-Known Member
'Stamets Stack' works for me.
Fangthane
Well-Known Member
And here's the new AIO bag I just got in the mail today. So dry that if you give it a little shake you can actually hear the pieces rattling around. I'm far from an experienced eye, but doesn't look to be anywhere near field cap to me. Just can't fucking win with these things lately.
canndo
Well-Known Member
I imagine that you could hear even moist pieces. You could inject 20 cc or so of sterile water
Drop That Sound
Well-Known Member
Your not supposed to shake up the AOI bags. They're supposed to stay in separate layers, at least I am pretty sure. Then you inoculate only the grain section. Once that layer colonizes, then you break up the bag and mix it all together.. right?
Fangthane
Well-Known Member
I'm not talking about violent shaking. Just moving it around, picking up or sitting down. The spawn is very loose and doesn't seem like much moisture providing surface tension to keep it together as I'm used to seeing with rye grain. In the bags that worked for me in the past, there was typically a decent amount of condensation droplets collected on the sides in the bag. This one looks much less moist than others I've used. I injected the last of my spores in it, but this is the first one where I've been this skeptical right off the bat and I don't have high hopes.