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jellyfish


Registered: 10/02/05
Posts: 7,457
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Need some advice UPDATED WITH PICS ADVICE APPRECIATED GREATLY 1
#23824055 - 11/11/16 08:20 PM (7 years, 2 months ago) |
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So I have recently got back into the hobby after a 3 year hiatus. I am going from spores from 3-1 years old onto malt extract agar in jars. I have a flowhood and did my best to be sterile. My goal was to immediately cut out healthy looking growth, but it all looks a bit fluffy. I did 4 jars each of ps. galindoi, ps. tampanesis, ps. mexicana and p. cubensis (golden teacher). It's been 2 weeks and most plates either have no growth or very fluffy white growth (although some stuff that's crept up the side of the jar looks like it could be rhizomorphic. I was experiencing some obvious non-white contams that I would make doing transfers easy, I did a control where I didn't inoculate and just opened them to air and they got nasty blue mould I'm going to nuke in my PC and then throw out.
The issue is these jars all have designs and stuff on them, showing a photo will be hard. I'm considering taking off the lids in front of the flowhood to take quick picture to get some opinions. I have never grown anything but cubes and I'm not sure how fluffy/w.e. the growth should be, it all looks like cobweb to me but I'm surprised that some jars showed no growth and some jars bright white growth, doesn't make it easy for me to know what part to cut out and transfer to new agar.
If you guys think it's a good idea, I'll sanitize the area, run the flow hood and take quick pictures to show you, ideally I'd like to transfer the healthiest portions to new agar plates. I feel like I might have mycellium and bright white mould and I'm unable (because of the designs on the jar) to know for sure) I see no yellow spots/myc piss indicating bacteria but some of these fluffy growths don't seem to merge with others. The growth all started from places where I had swapped spores but the prints could have not been the cleanest. The cube print I took 3 years ago.
So should I wait for the plates to fully colonize? Or would you advise that I open them up under a flow of laminar HEPA filter purified air and post pictures? I feel maybe these are all failures, but in my younger days, when I messed up, I would see all sorts of mould. Everything I see is bright white, just different morphologies. I have time Sunday to prepare more plates and do transfers I'm just not sure if I'm just cultivating some kind of white mould.
If you check my sig, you can see my flowhood and agar plates. The fluffiness makes me think cobweb but it's hard to take a photo through the patterns in the glass.
Any help would be greatly appreciated.
Edited by jellyfish (02/25/17 10:48 AM)
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PirateSwazey


Registered: 12/12/12
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Re: Need some advice [Re: jellyfish]
#23824095 - 11/11/16 08:40 PM (7 years, 2 months ago) |
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I would take a picture after you do your next transfers.
It sounds like you have cube myc, it's not uncommon at all for growth from spores to be mostly tomentose. Make a few rapid transfers and look for rhizomorphs, then transfer from those a couple more times.
As far as working with the jars go, why not order some plastic petris and make it easier on yourself in a few weeks?
Since you got mold on your control plate, when is the last time you changed your pre filter on your flowhood?
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A.RichardTrickle
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Registered: 11/04/16
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I concur, I have used MS for most of my experiments over the past few years and found new growth from spores is almost always fluffy. Best wishes Dick
-------------------- "When eating shit, it is best not to nibble. Bite, Chew. Swallow. Repeat." "If you're making love to your old lady, someone else is fucking her" "Douchebags are children who never grew up, like Sheeklette, we should pity them." [quote]Niffla said: [quote]A.RichardTrickle said: Dick[/quote] http://www.youtube.com/v/kbwNUOUy-3c[/quote]
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jellyfish


Registered: 10/02/05
Posts: 7,457
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I opened the control in open air, not in front of flow hood. The jars that show no growth at all (despite having spores swabbed on them, were opened in front of the flow hood which makes me think it's working fine. The prefilter is just a bunch of cheese cloth over the top of the fan. I read some rule of thumb that the flow should only be strong enough to make the flame on a lighter bend but not extinguish but I don't remember where I read it and if it was supposed to bend at a 45 or 90 degree angle. I've never changed the prefilter but I also haven't ever used the flow hood too many times in total. Probably less than 20/30 times.
edit: no point in waiting till full colonization for transfers? Also, can anyone let me know what I should be looking for for the truffle forming species? I know how to pic out nice rhizomorphic cubensis growth but this is my first time with truffles. On Sunday I will make a new batch of agar and then take photos and do transfers (can't find my scalpel damn it). I'm guessing it makes no sense to let it fully colonize. I am just unsure in this bright fluffy white mess what sections would be best to transfer and what good tamp/mexicana/galindoi myc looks like. I was under the impression it was more greyish and also, it looks kind of clumpy like clumpy white fluff. I don't use petri dishes because I find it easier to PC the agar already in the jar. I should just get clear regular jars. Thanks a lot guys. All input is appreciated.
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Grundalizer
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Re: Need some advice [Re: jellyfish]
#23824148 - 11/11/16 09:10 PM (7 years, 2 months ago) |
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Any white mycelia, fluffy or not, which is derived from a known spore addition to a sterile media under sterile or semi-sterile conditions and is growing out of the area you doped the media with, is 99.99% what you are looking for. Cobwebby growth does not equal contamination. I'm a microbiologist and mycologist by trade and I'm amazed at how often I see people on hear freaking out about contamination and throwing things out (not trying to call you out, I just don't post much, and am trying to start to more). If you have anything growing on a plate, just pick a corner with a sterile scalpel or sterile toothpick and re-plate...subculture till purity.
Any moulds are easy to see because of their almost instantaneous rapid color changes due to conidia with spores forming which change color often due to their age.
Any true basidiomycete will stay in a white mycelial growth, and yes, often if I regenerate old spores or super old dried our desicated agar plates, initial growth is very very very wispy.
I found sub-culturing doesn't help, but taking some and dropping it into sterile broth, like potatoe dextrose broth, yields very healthy and dense mycelial growth...which I then re-plate onto solid agar.
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PirateSwazey


Registered: 12/12/12
Posts: 2,993
Loc: Here, Now
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The tamp mycelium will look fainter and stringier than what you're used to with cube myc.
Sounds like your flowhood is working fine... 45-90 degrees should be on.
Yes it's perfectly fine to transfer before the plate grows all the way out.
Sorry I just don't get why you have a flowhood but do no pour agar?
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Basically I started as a newb with pf tek. Now I have a punch of jars, with lids on them that have 4 holes poked in them for pf tek style inocs, that I've converted to tyvek lids. So everything I use is stuff I've converted via tyvek from newb times before I built my flowhood. I don't have any petri dishes, maybe 2, I remember I broke one. I guess I should get petri dishes. I just like for whatever reason, pressure cooking jars with tyvek lids and then inoculating them in front of the flow hood. That way if something goes wrong, it only goes wrong with one plate as opposed to if I poured the molten agar in front of the flow hood and something went wrong, I would ruin the whole batch.
I can't tell you guys enough how grateful I am for the fast responses. I joined this site in 2005 boiling pf tek jars and inoculating in my kitchen with a syringe. I finally built a flow hood in 2012 then got distracted with life and now I'm back and trying to figure shit out.
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PirateSwazey


Registered: 12/12/12
Posts: 2,993
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Re: Need some advice [Re: jellyfish]
#23824360 - 11/11/16 10:42 PM (7 years, 2 months ago) |
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Hey it's just matter of preference.
Pouring plates is easy in front of a hood though.
Best luck man!
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jellyfish


Registered: 10/02/05
Posts: 7,457
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I'll be back with pics soon because honestly I'm rusty. I went from pf tek to pf tek with casing to really shitty glove box with rye which I could never spawn successfully. I only had 2 grows with my flowhood, so my experience with agar is limited. Since I built the flow hood my grows have been amazing, no spore syringes, just prints, no contams at all and spawning to bulk was a piece of cake as well but it's been 3 years since I went from print, to grain, to coir/verm and I have forgotten a lot. I'll probably post pics sunday, and make a few transfers and hopefully I can get some more input then. I need to see how much tyvek I have left, the stores near me don't sell those tyvek paint suits anymore. And jars, I want to make as many agar plates Sunday as possible. Thanks again everyone. I'll be back with pics/news. I will be very generous with prints if this all works out and I can generate some.
edit: one more question, the plates that I swabbed over 2 weeks ago that still show no growth, I don't exactly have a ton of tyvek lids/half pint jars, should I nuke them along with the contamed plates or is it possible I will see growth after 2 weeks. I need to make new agar jars tomorrow and to do so, I need to at least free up some of the jars I have, so I am going to nuke the 2 contamed jars (out of good practice to kill any potentially dangerous mould), then I'm going to make as many agar jars as I can, do the transfers and take pics while I do them.
Edited by jellyfish (11/12/16 11:45 AM)
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Big Bear
Earf Child



Registered: 06/11/14
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Re: Need some advice [Re: jellyfish]
#23825930 - 11/12/16 01:23 PM (7 years, 2 months ago) |
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I just had some wikidzons take a little longer than 2 weeks to germ. I'd say say get a sleeve of petris or more no pour plates rather than having to clear cut for new upscale mycelium luxury condos.
I fucked myself by not having enough plates earlier in the year, I took some mold with me somewhere along the line, and then everything got fucked. These days when I take transfers and open plates and start spores I go crazy with it and make way more than I need.
I like petris a lot. I do better with them personally. Just do yourself the favor and get a 500 pack for ~ 100 bucks and it'll last you a really long time.
The User bodhisatta has a really good writup on agar, that I pretty much go by. Only difference is I use parafilm cuz I like it
Good luck
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Re: Need some advice [Re: Big Bear]
#23830255 - 11/13/16 07:35 PM (7 years, 2 months ago) |
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Thanks a lot. I'm taking a look at bodhisattas tek now. I have a nice 500 mL erlenmeyer flask and I'm going to buy some petri dishes. I found 2 at home today so I just put the agar in them first and PC'd them, I know it's not the best way to do it but hopefully I can get some growth.
I don't know why I thought I'd have pics to show you all today. I did all the agar stuff hours ago and the jars are still warm and the agar is still molten (last I checked anyways). The person I live with recently got over lung cancer and I didn't want to open any contam'd jars so I had to run the PC once first just to eradicate the contams before cleaning the jar out to reuse it. Now I have about 16 jars/plates in total so once they are cool I'm going to open up my cultures, take photos and do transfers. I couldn't find my scalpel blade so I pressure cooked my nail clippers (because they have a little knife thing on them and they're metal), and a pair of tweezers.
Is 70% IPA enough to clean the tweezers/knife between transfers in front of my flow hood or should I get 99%?
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Big Bear
Earf Child



Registered: 06/11/14
Posts: 5,415
Loc: In love, On time
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Re: Need some advice [Re: jellyfish]
#23830371 - 11/13/16 08:10 PM (7 years, 2 months ago) |
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70 is good
Edit: didn't read well. I thought u meant normal sanitation for which 70 is fine. Flame for transfers
Edited by Big Bear (11/14/16 12:51 AM)
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
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Loc: Canada
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Re: Need some advice [Re: Big Bear] 1
#23830844 - 11/13/16 11:11 PM (7 years, 2 months ago) |
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You should be flaming tools in between transfers. . .
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PirateSwazey


Registered: 12/12/12
Posts: 2,993
Loc: Here, Now
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Only time you don't need to is when you're using multiple wedges from the same plate to inoculate the same grain master.
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Yea I flame my inoc loop on my alcohol lamp I just thought maybe the knife and tweezers are a bit too thick of metal for me to see that nice red glow that let's me know they've been under the flame long enough. I haven't posted the pic in this thread but in the journal linked in my sig there's a pic of my flow hood with my alcohol lamp burning which I use for flaming.
Edit: when I was an undergraduate student we would dip our innoc loop in alcohol then flame it. I kind of forgot about that technique and I'm afraid of causing a fire but should I do it that way or just hold tools over my flame
Edited by jellyfish (11/14/16 07:45 AM)
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PirateSwazey


Registered: 12/12/12
Posts: 2,993
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Re: Need some advice [Re: jellyfish]
#23831469 - 11/14/16 08:17 AM (7 years, 2 months ago) |
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Buy a scalpel with disposable blades. The blades will be thin and turn red very quick. I like the #7 handle with #11 blades. Very cheap on eBay!
If you like to dip your tools in alcohol before you flame them it isn't going to hurt anything. Guessing they taught that in your classes to clean off any soot or agar that may get on there.
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jellyfish


Registered: 10/02/05
Posts: 7,457
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I figured it was because it would burst into flame which was a visual indicator that they had gotten hot enough. I modified my scalpel to hold a razor blade for taking cuttings of cacti and now I can't find the original scalpel blade so I will get a new one. The growth in my dishes looks a lot healthier than I thought and I think I even see some rhizomorphic growth so the only issue now is that I'm not sure how to select healthy mycellium from the non cubensis species I'm growing. I have about 16 agar plates/ jars to use tonight and like 12 jars of healthy looking mycellium of tamp, cubes, and mexicana. My galindoi jars still show no growth. I really appreciate all the advice and will have some pics coming soon but I'm actually quite hopeful. Building a flowhood was the best move I ever made in terms of this hobby. Contams have become a rarity. It cost me $500 CAD to build the flowhood and the confidence it gives me in terms of successful inoculations makes it worth every penny.
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PirateSwazey


Registered: 12/12/12
Posts: 2,993
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Re: Need some advice [Re: jellyfish] 1
#23831667 - 11/14/16 09:38 AM (7 years, 2 months ago) |
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Quote:
Building a flowhood was the best move I ever made in terms of this hobby. Contams have become a rarity
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Didn't have time for transfers yesterday. Hydro store nearme sells 100 plastic petri dishes for 30 bucks which I assume can't be pressureduced cooked. also have expensive and in my experience, very dilute spores syringes and bags of Rye and brown rice flour yoi could get at bulk store down the street for 10x cheaper but it's cool they're catering to the noobs for the hobby. I bought a presterilied scalpel but I don't wanna open it but I feel the handle might be plastic therefore I can't pc it either.gonna try to do transfers and post pics tonight if I get home early enough from work.
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spore-ty



Registered: 01/21/16
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Re: Need some advice [Re: jellyfish]
#23835121 - 11/15/16 11:09 AM (7 years, 2 months ago) |
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Dam I wish my local hydro stores hooked us up
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Grundalizer
Stranger

Registered: 03/15/15
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Re: Need some advice [Re: spore-ty]
#23836633 - 11/15/16 07:25 PM (7 years, 2 months ago) |
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I can get you a case of 500 gamma-irradiated sterile petri dishes in sleeves of 20 for $83.
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A.RichardTrickle
Feel like a Stranger

Registered: 11/04/16
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Quote:
Grundalizer said: I can get you a case of 500 gamma-irradiated sterile petri dishes in sleeves of 20 for $83.
Someone want to split it with me?
-------------------- "When eating shit, it is best not to nibble. Bite, Chew. Swallow. Repeat." "If you're making love to your old lady, someone else is fucking her" "Douchebags are children who never grew up, like Sheeklette, we should pity them." [quote]Niffla said: [quote]A.RichardTrickle said: Dick[/quote] http://www.youtube.com/v/kbwNUOUy-3c[/quote]
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Those petri dishes sound tempting.
My agar jars before germination:

Growth from my 3 year old spore print. Can't tell if it's cubensis or cobweb mould or something. The bit of the left that looks a bit rhizomorphic was removed yesterday and transferred to fresh agar. What do you guys think? I took pictures of the growth on my mexicana and tampanensis plates but the lighting was awful and they didn't work out. I didn't know how to choose what parts of the agar to transfer. Do you look for rhizomorphic growth like you do with cubes?

My flowhood is the coolest thing I've ever built myself
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enlightenment
alchemist


Registered: 08/09/09
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Re: Need some advice [Re: jellyfish]
#23840509 - 11/17/16 12:57 AM (7 years, 2 months ago) |
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Looks good  If you have a flow hood i highly recommend to use petri dishes. They need less space and it is easier to take a look at the growth. They can not be pre poured but with a flow hood that shouldn't be an issue.

A disadvantage is that petri dishes can not be reused unless you buy glass petri dishes. I had glass dishes once but sold them because I hate to clean them.
Edited by enlightenment (11/17/16 01:03 AM)
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jellyfish


Registered: 10/02/05
Posts: 7,457
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See I have always done things a little more simply, like spore syringes or taking flesh from a healthy mushroom. But now that I'm trying to go from spore prints, and contams are inevitable, I'm really seeing how crappy jars are. Ideally I should be able to glance at my agar and immediately see the whole 2D picture of what's going on. I should be able to spot contams right away. Also I have no idea what I'm doing with the sclerotia forming species so it would be a lot easier for me to show you all pictures and get feedback. I'm going to have to buy some petri dishes, I just don't like disposable stuff, I want glass ones I can reuse.
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enlightenment
alchemist


Registered: 08/09/09
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Re: Need some advice [Re: jellyfish]
#23840558 - 11/17/16 01:42 AM (7 years, 2 months ago) |
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Quote:
I just don't like disposable stuff, I want glass ones I can reuse.
Good attitude. 
If you don't want to do a lot of species at the same time, using glass dishes is a great idea. Good luck
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wtfcrazymofo
foil hater



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Re: Need some advice [Re: jellyfish]
#23840636 - 11/17/16 03:22 AM (7 years, 2 months ago) |
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-------------------- If you want to eat->https://www.shroomery.org/forums/showflat.php/Number/8553541 Bag sealers are to bulky (my hood isn't that big) https://www.shroomery.org/forums/showflat.php/Number/28622922
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Hey guys. one of my new agar plates from a trasfer I did (cubensis) is showing amazing rhizomorphic growth. Only issue is a bit farther away from the agar wedge is a white spot that doesn't look rhizomorphic. Now, it could be that I was sloppy when I added the agar and a tiny piece broke off, but I'm afraid it's a contam. I want to transfer the super nice growth asap to a new agar plate, but I won't have time until at least tomorrow night, possibly even a few nights. I decided to put it in a ziploc bag and put it in the fridge to slow down the eventual collision between my nice rhizomorphhic splendor and this possible white mould. Bad idea? I'm going to try to make new agar plates tomorrow but honestly I have so many people I live with who are always cooking it's rare for me to get a stove top to do my stuff on.
Hope the fridge wasn't a dumb idea. I put it in a clean ziploc just because I know fridges are notorious for contams.
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A.RichardTrickle
Feel like a Stranger

Registered: 11/04/16
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Re: Need some advice [Re: jellyfish]
#23852740 - 11/20/16 09:14 PM (7 years, 2 months ago) |
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Pic?
-------------------- "When eating shit, it is best not to nibble. Bite, Chew. Swallow. Repeat." "If you're making love to your old lady, someone else is fucking her" "Douchebags are children who never grew up, like Sheeklette, we should pity them." [quote]Niffla said: [quote]A.RichardTrickle said: Dick[/quote] http://www.youtube.com/v/kbwNUOUy-3c[/quote]
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Can't get a nice pic through the jar but on one side of the plate there's an agar wedge with beautiful rhizomorphic growth spreading out from all sides and on the other side is a bright white spec I'm afraid is a contam. If I don't have time to make agar tomorrow then transfer has to wait till next Monday cause I'm too busy. Was fridge a bad idea?
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,810
Loc: Canada
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Re: Need some advice [Re: jellyfish]
#23852923 - 11/20/16 10:56 PM (7 years, 2 months ago) |
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The fridge should be fine. It will slow everything right down.
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jellyfish


Registered: 10/02/05
Posts: 7,457
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So I seem to have isolated some rhizomorphic growth from my cubensis spores. The galindoi and tampanensis spores (a year old) didn't germinate. I have isolated some of what looked like healthy growth from each of my 4 tampanensis "plates" to 2 new plates each. Mycellium still seems really fluffy. Until I have a lamp by my flowhood I will not be able to provide good pics, but I think with one more transfer I will have a really nice cube culture. Can anyone direct me to pics of what sclerotia forming species are supposed to look like on agar? I am so used to cubes.
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PirateSwazey


Registered: 12/12/12
Posts: 2,993
Loc: Here, Now
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Re: Need some advice [Re: jellyfish]
#23877879 - 11/29/16 10:39 AM (7 years, 2 months ago) |
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Thanks a lot that's really helpful
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jellyfish


Registered: 10/02/05
Posts: 7,457
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So I will call the spores to agar plates gen 1. For cubensis (golden teacher spores I took myself 3 over years ago) gen 1 was pretty dirty. Some gen 2 plates are contam'd but a couple have nothing by mycorhizzal goodness. Because it's christmas baking season I'm not sure when I'll be able to PC some rye so I'm thinking of putting the best cultures in ziploc bags in the fridge and occasionally taking them out to open them to let them breathe as I know even with tyvek, fridges are full of mould spores. Speaking of tyvek the 2 stores that sold tyvek painting suits are gone from my area so I need to find a new source. One of the best looking plate actually fully colonized and then fruited a sad little shroom. For gen 3, I took all the best rhizomorphic growth, except for one plate where I just extracted the fruting body and put him on directly. Looking back that may have been done, should have cut the cap off so I don't have spores released cause I'd like some type of isolate. Every other gen 3 plate came from a clean gen 2 plate except the one I had put in the firdge that had brilliant rhizomorphic growth but contam on the other side of the plate, which I sliced out last night for a gen 3 culture. If my gen 3 cultures wen't well then I have about 3 fully colonized rhizomorphic gen 2 plates, (one of which fruited), and 5 gen 3 plates I tried my best to isolate from the nicest growth. Like I said, I don't want my gen 2 plates fruiting again so is fridge the way to go until I have time to get some rye going?
Tampanenesis and galindoi prints I bought from a respected vendor did nothing, I have growth on the mexicana plates. Gen 1 looked nasty. Gen 2 looked a lot less nasty but still some obvious contam. Did my best to isolate the mycellium from gen 3 that looked like the pictures posted above. I am wondering, like I said it will be a while until I have rye going. The gen 2 plates that are clean, I may refrigerate so they don't end up fruiting in the jar. I made the agar thick so that they won't dry out too quick but they are fully coloznized with beautiful perfect looking growth. The gen 3 plates I made still have a week or two to colonize. My question is, should I leave the mexicana plates (gen 3 which is hopefully clean, and gen 2, a couple of which don't look contamed, and wait until I see sclerotia form? Then transfer those to rye in order to be sure I have the right stuff? This will buy me some time until I can get rye going. The mycellium bruises blue when I cut it which is a plus. Still don't have a scalpel so I pressure cook my whole nail clippers in a tyvek lidded jar and use the little knife attached to them and flame it every time over my alcohol lamp.
Absolutely loving this flow hood. My basement is beyond dusty. Food moulds so quick, we even had a mould issue on the walls at one point. Yet the majority of my cultures are clean. Wish lighting in my basement was better so I could show you the mexicana agar cultures but the cubensis are picture perfect atm. 3 year old spore print, first spore print I've ever taken. Pretty awesome.
edit: Don't want to double post but to sum up what I'm asking:
1) should my fully colonized plates cubensis plates with beautiful rhizomorphic growth be okay in the fridge in ziploc for a bit (couple weeks) before I am ready for rye? I will take them out periodically and open them to let fresh air in. I don't want them fruiting on agar.
2 a) For the mexicana plates that seem okay, should I wait to see sclerotia before moving to rye
2 b) Or should I wait to see sclerotia and then transfer that to fresh agar, as I know it's prone to truffle formation? Thanks.
All these are half pint jars with agar and tyvek lids I made almost a decade ago. I need new tyvek but haven't had the time to research who still sells those suits. I absolutely love tyvek + flow hood. Makes this hobby a breeze. I can open sterile agar in front of my fan in the dustiest basement for 5 minutes and get no contams. I just got a scalpel too which I can PC before use and then flame with my make shift alcohol lamp.
Next step is to grow something legal. Only growing the cubes to get prints as my prints were 3 years old and wanted to revive them before it was too late. Also gives me something to trade with you guys.
Edited by jellyfish (12/07/16 11:36 AM)
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jellyfish


Registered: 10/02/05
Posts: 7,457
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So pretty much every one of my cultures is nothing but mould. Dunno what I did the last set of transfers. Bleached the area, worked in front of laminar flow hood, flamed tools in between uses. Even pressure cooked the tools before use. But now I have to start from scratch. There's a chance one or two of my nice cubensis cultures are clean but all my mexicana are nothing but mould. I've never had this happen since getting my flow hood. So sad.
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Alright I'm sick of this tyvek jar thing. How do these petri dishes come? Are they all individually sealed? I have a flow hood but even still I'm worried about contam. Also all my golden teacher jars show rhizomorphic growth I'm just stalling till holidays are over since I can't do any rye work while the kitchen is so busy.
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jellyfish


Registered: 10/02/05
Posts: 7,457
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So I am still unsure if I am growing mexicana or some sort of wispy white mould since I've never worked with it before. I did pluck this out of one of my agar jars. A piece of scleortia perhaps?
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Re: Need some advice [Re: jellyfish]
#24054921 - 01/31/17 03:39 PM (6 years, 11 months ago) |
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Just an update. Managed to revive 3 year old GT spore print I took myself as well as a mexicana spore print I bought years ago. The other sclerotia forming spore prints I bought at the same time gave me nothing but mould and isolating a clean culture from the mexicana was a bit difficult. Wasn't used to the appearance of the mycellium and had a lot of mould initially but finally cleaned it up.

The leftmost 3 jars are mexicana, the last 2 are golden teacher. My agar cultures are drying out and the GT agar jars are all fruiting off the agar. I am assuming I should just ditch them, clean them and then use these rye jars to inoculate more rye and also start more agar cultures. I have read different things about whether or not to shake mexicana rye jars. Also, I am planning on using one of the three mexicana jars for G2G to start more rye. This should be fine once the jar is fully colonized correct?
Edited by jellyfish (01/31/17 03:45 PM)
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Mikeify
Wildin Foo



Registered: 01/30/17
Posts: 288
Loc: USA
Last seen: 3 years, 2 months
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Re: Need some advice [Re: jellyfish]
#24055302 - 01/31/17 06:09 PM (6 years, 11 months ago) |
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Im also looking forward to growing GT's but are they prone to bacteria?
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Re: Need some advice [Re: Mikeify]
#24055367 - 01/31/17 06:31 PM (6 years, 11 months ago) |
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I'm using GT as an abbreviation for golden teacher. Since a cube is a cube, I don't see why they would be more prone to bacterial infection than any other variety of cubensis. The print I used to get that rye going was taken by me from my first and only monotub over 3 years ago. I got several flushes out of it and would re-hydrate the bulk sub by taking the tub into the bathroom and filling it up with a bit of water from the bathtub faucet. I wanted to scrap it before it got contaminated, so I did. Had no issues with bacteria.
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jellyfish


Registered: 10/02/05
Posts: 7,457
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Re: Need some advice [Re: jellyfish]
#24118539 - 02/25/17 10:47 AM (6 years, 10 months ago) |
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Don't want to straight a new thread but need advice. I thought I had finally isolated mexicana and golden teacher on afar over successive transfers. One transfer even involved using a stone that formed on a mexicana plate. I inoculated rye, but didn't get a chance to inoculate for a week or so and I'm afraid it may have dried a bit. Also shook all my jars to speed colonization.
I have one jar of mexicana and one jar of GT that seem alright. All my agar cultures I'm going to give up on and start over because they have gotten dry and don't seem too successful. If these 2 jars I post seem okay, I will use them to start a new set of agar plates of mexicana and GT as well as a few spores I got through some trades. My main goal is to grow sclerotia as I have heard mycellium is legal in Canada just not fruiting bodies.
The ones that seem okay: Mexicana:
 GT

There are also 2 other mex jars and 1 other GT jar. They all look like they didn't colonize fully, either I introduced bacteria by shaking, they stalled, or they're something else entirely. It was hard to get a clear pic but one of the mex jars has a thick layer on top that looks more like ice than mycellium of any sort.
Weird mexicana jar1 (with that thick ice looking growth




Weird mexicana jar2:
 messed up this pic. Meant to show how some spots at the bottom are uncolonized and they were inoculated a month ago. Good be the agar got too dry but I feel it's bacterial. I use a flow hood and pressure cook my rye for over an hour so maybe shaking somehow got something through the tyvek lid.
Weird GT jar 1:

Probably going to ditch all but the 2 okay looking jars, use sterilized tools in front of a flow hood to try to inoculate some agar while still letting the mex jar be able to slowly form stores.
Advice very appreciated.
I feel I might need to make this a new post because people are going to see how many replies this thread has and assume my questions have been answered.
Most of my spore collection is over 3 years old. The GT print I took in 2013 actually fruited on agar but obviously one of my 2 GT jars are either stalled or contamed. Should I bother transferring the two decent looking rye cultures back to agar and maybe start some new spores and leave the rest of the mexicana jar to see if it will produce stones. I used jars with holes in the lids like I would normally with cubensis on rye. Should the holes be smaller to encourage CO2 buildup and cause stones to form?
Thanks again.
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