Becca's B+, Blue Meanie, Purple Mystic From Spore Genetics & Blue Yeti LC Fun Run

[BeccaBuds]

The deed is done! 5 jars of liquid culture, 3 Uncle Ben's Rice Bags and one Midwest Grow Kits All-in-One (AIO) bag have been inoculated this evening. Tarp over SAB was effective but a major PITA, but ya gotta do what ya gotta do.

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Here's the inoculation line up:

• Liquid Culture (LC) jar 1: B+ spores​
• LC jar 2: Blue Meanie Cubes spores​
• LC jar 3, Rice bag 1 & 5: Purple Mystic spores​
• LC jar 4, Rice bag 2 & AIO bag: Blue Yeti liquid culture​

Rice bag 5 is the bag I nuked for 90 seconds in the microwave so it will be interesting to see if there is a difference in growth compared to bag 1. The AIO and Rice bag to will be interesting as they are different grains.

In retrospect: I think I forgot to flame sterilize the needle for the Blue Meanies, though I did wipe the SHIP and the needle with an alcohol wipe, so I hope the LC doesn't get contaminated. When I autoclaved the liquid culture jars I forgot to cover them with foil but I did make sure the lids were tight beforehand. Now the fun begins (knock on wood)!

 

[Stoned Ape]

Game on

 

[BeccaBuds]

One final thing to do tonight, then I can clean up and kick it; got out the new mag stirrer and gave each jar of LC a whorl to mix up the B+, Blue Meanie & Purple Mystic spores & Blue Yeti LC. That gets done again in 36 to 48 hours (Monday). Everything inoculated with the Blue Yeti culture should get cooking pretty fast as it's already an isolated dikaryotic mycelium and ready to multiply. What I've read says like a week faster at least.
Regarding the liquid culture premix: I got a jar of dry mix from amazon by a company name Yuantide. It contains dextrose, Light Malt Extract, Soy Peptone and bakers yeast, though I've read a lot of people use just Karo or honey. They say Karo and highly filtered light colored honey are easier to observe the growth of the culture because they are nearly colorless in solution. I may try that later but we'll see how this goes.
I should be able to put them in the fridge for long term storage in a week or 2.

 

[BeccaBuds]

Somehow missed listing LC jar 7 in my earlier post tabulating inoculations. It is a smaller 200mL jar of LC inoculated with Blue Yeti LC along with jar 4, rice bag 2 and the AIO. I can see clumps of mycelium in both jars 4 & 7 already so life, I'm sure, is getting a foothold in rice bag 2 and the AIO as well!

 

[BeccaBuds]

Did the 48 hour agitation of the 5 liquid culture jars with the mag stirrer after taking photos. No sign of life in the 3 injected with spores yet? The jars injected with liquid culture are going full steam ahead. Check jars 1-3 again in 2 days. No sign of life in the rice bags or AIO yet either.

 

[BeccaBuds]

I think I am seeing signs of life in jars 1-3 (spores inoculated) now but still very minimal. Jars 4 & 7, inoculated with liquid culture, both have huge colonies forming, covering the whole bottom of the jars 10mm deep. Saw an odd spot on the filter disk on jar 1 (B+ spores) that I hope isn't contamination. Spritzed it with 70% iso just in case.
I'm going to get some pre-poured petri dishes for testing the LC jars so I can verify them before inoculating any grain.
Ouch, another $30 for the petri dishes & some polyfilm tape. A step I hadn't considered, but necessary using liquid cultures because you can't fully identify what's growing in the liquid by sight alone.
No sign of mycelium in the grain bags in the SAB yet. Temp in the SAB is 76F at 84% RH. Opened the FAE ports and spritzed the filters with 3% hydrogen peroxide, which will decay to oxygen and water to raise the RH a bit. MGK says it won't hurt their filter disks that they get from Microppose. Closed the ports back up again.

 

[BeccaBuds]

Just stirred all the jars of LC again today. This mag stirrer is really cool- when you crank the speed up a bit it churns the air above the liquid into the liquid. That should help 'em breathe! I can definitely see some action in jars 1, 2 & 3 now (B+, Blue Meanie & Purple Mystic spores, respectively). Not much but there's life in there of some kind forming. Jars 4 & 7 (Blue Yeti LC) continue to amaze and both are near 1/4 colonized now. Rice and AIO still no sure signs of mycelium. This weekend I'll open the SAB and check the bottoms of the Blue Yeti rice and AIO bags for growth.

Jar 7 before stirring today. That dark mass at the bottom is the stir bar obscured by the mycelium.​

 

[BeccaBuds]

Day 7 update. Time for the mag stirrer again in all the liquid culture jars to mix and oxygenate the liquid. There is life visible in all three spore inoculated jars. The jars inoculated with liquid culture are almost ready for the fridge and culture identification when the pre-poured petri dishes arrive. The structure of the masses in jars 1 & 2 look similar to 4 & 7 while Jar 3, the Purple Mystic, formed a bunch of separate BB sized masses.
Nothing to see yet in any of the grain bags yet so I'll let them be for now. Temp and humidity in the SAB is 78F at 85%RH.

 

[Absorber]

Day 7 update. Time for the mag stirrer again in all the liquid culture jars to mix and oxygenate the liquid. There is life visible in all three spore inoculated jars. The jars inoculated with liquid culture are almost ready for the fridge and culture identification when the pre-poured petri dishes arrive. The structure of the masses in jars 1 & 2 look similar to 4 & 7 while Jar 3, the Purple Mystic, formed a bunch of separate BB sized masses.
Nothing to see yet in any of the grain bags yet so I'll let them be for now. Temp and humidity in the SAB is 78F at 85%RH.

This is all totally foreign to me I've never grown or had psychedelic mushrooms.
Have you got multiple mag stirrers that you leave in the cups so you don't have to fiddle with getting them out or making sure there's no cross contamination.
Good luck with it all

 

[BeccaBuds]

This is all totally foreign to me I've never grown or had psychedelic mushrooms.
Have you got multiple mag stirrers that you leave in the cups so you don't have to fiddle with getting them out or making sure there's no cross contamination.
Good luck with it all

@Absorber You put a little bar magnet (coated with teflon plastic) in each jar with the liquid. The stir table has a magnet with a variable speed motor to spin it that couples magnetically to the bar in the jar to spin it without contact. No need to even open the jar! I got this one off Amazon for under $30 plus $10 for the stir bars 10-pack on the left.

 

[Absorber]

@Absorber You put a little bar magnet (coated with teflon plastic) in each jar with the liquid. The stir table has a magnet with a variable speed motor to spin it that couples magnetically to the bar in the jar to spin it without contact. No need to even open the jar! I got this one off Amazon for under $30 plus $10 for the stir bars on the left.

Thanks , I've seen them before I just wasn't sure if you had a magnet stirrer for each jar ,but it looks like you have .
Thanks for the explanation though and the reply

 

[BeccaBuds]

Day 9 since inoculation. Time to stir the LC jars again. Jars 4 & 7 look like they will be ready to move to the fridge tomorrow and the other 3 jars aren't more than a day or 2 from being fully colonized as well. It appears the trick is to stir them fast enough to draw in air as well at stir them up. Growth can really explode when more oxygen is mixed into the medium.

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Still no visible growth in the AIO but I think that's because I shot the LC deep into the bag instead of close to the surface like the photo in the instructions they ship with it. I am confident I'll see mycelium in it soon. When that happens I'll crack the SAB (wearing clean clothes with sterilized gloves and a surgical mask) for a closer inspection of both rice bags and AIO.

 

[BeccaBuds]

Day 11 after inoculation. Some changes in the condensation on the inside of the AIO bag but nothing I can be certain of. Instructions say 25 to 45 days for full colonization depending on strain so I'm putting off hand inspection until Saturday. Gave each of the jars another vigorous stir. There is a concerning dark speck floating around in jar 3, the Purple Mystic spores, see the following post. Here are pictures taken before and after stirring:

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[BeccaBuds]

The speck floating in the Purple Mystic jar (upper middle below the bubbles). If it's bigger tomorrow, I'll put it in the fridge until the petri dishes arrive to test the culture.

 

[Lady Cannafan]

The speck floating in the Purple Mystic jar (upper middle below the bubbles). If it's bigger tomorrow, I'll put it in the fridge until the petri dishes arrive to test the culture.

That speck would be hopefully mycelium? I am totally unfamiliar with this method, but today was a plan to do some liquid culture beginnings.

 

[BeccaBuds]

No, @Lady Cannafan! The mycelium is the cloudy looking stuff. The black speck is either some piece of harmless junk in the jar or unknown biohazard to the mycelium. If it gets no larger it's probably not biological. I'll have to sample the culture and grow it out on agar in petri dishes and see what grows.
It didn't break up with vigorous stirring so I'm hoping it's a piece of junk that slipped through, that was sterilized when I autoclaved the jars.
I did the cultures right off the bat so I would have reserves in case I screwed anything up. Plus LCs colonize quicker.
If you haven't seen Willy Myco's vids on youtube regarding Master Slurry cultures, I highly recommend it. You may want to try that instead. It eliminates the need to test the culture on agar since you're starting on agar to begin with and contamination becomes obvious.

 

[Lady Cannafan]

Thanks Becca!

 

[BeccaBuds]

I just inspected jar 3 again this morning and the speck seems to have disappeared. I may have an explanation but continued observation needed...
When I removed the jars from the pressure cooker, I noted brownish-black sludge had precipitated to the bottom of the jars and covered the white stir bars. I believe it to be one of the ingredients of the commercial LC pre-mix powder I used, possibly nutritional yeast or peptone. I'm theorizing some of it broke off the stir bar during stirring and the mycelium munched it. Or so I hope!
I'll check it again tomorrow when I give them another stir. Jar 4 is almost fully colonized, less than a half inch to get to the surface of the liquid. Jar 7 still needs a bit longer before storing in the fridge, as do 1-3.
Still no visible white stuff in the AIO...

 

[BeccaBuds]

Just realized I forgot to order a roll of PolyFilm tape from Microppose. Missed their Black Friday sale. Darn, everything was marked down 40%, sigh. Have to have it though for the Rapid Rhizo petri dishes coming from Midwest Grow Kits so click, click, ka-ching! and the supplies are on the way.
Also got a card of (50) self-adhesive 0.22um, 20mm diameter jar filters. The ones I got off Amazon all look a bit bulged up after pressure cooking, like they might not have adequate FAE, so I want to try the Microppose filters. There were a few bad reviews on the Amazon filters, but all good for the Microppose on their website and a few youtube channels that mentioned them.

 

[Lady Cannafan]

Just realized I forgot to order a roll of PolyFilm tape from Microppose. Missed their Black Friday sale. Darn, everything was marked down 40%, sigh. Have to have it though for the Rapid Rhizo petri dishes coming from Midwest Grow Kits so click, click, ka-ching! and the supplies are on the way.
Also got a card of (50) self-adhesive 0.22um, 20mm diameter jar filters. The ones I got off Amazon all look a bit bulged up after pressure cooking, like they might not have adequate FAE, so I want to try the Microppose filters. There were a few bad reviews on the Amazon filters, but all good for the Microppose on their website and a few youtube channels that mentioned them.

I hope you're happy. You caused me to spend more money today on supplies with that post!
I can't afford following you. I keep getting reminded of what I'm going to need...