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OfflineEntheoGod
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Help me find my error in WBS prep?
    #26785017 - 06/26/20 03:00 PM (3 years, 6 months ago)

I have been needing to make this a post for a while now but have been busy failing. 😅 So to start this off let me explain my situation. I have been doing this for 5 years and have never had so much trouble getting something, anything, growing. I have the patience and am willing to put in the time for anything needed. Recently even started agar because of the lack of success I have been having with straight MS syringe to WBS jars. I have been preparing my seeds in several different ways each time just to see what I could be doing wrong. I have tried 24 hour soaks with no simmer, 24 hour soaks WITH a simmer (200F water right before a boil for 10-15 minutes.), 60 minute hot soaks 200F water no simmer, 15 hour soaks and on simmer, etc.. For the past 4 or 5 months I havent been able to get a single thing to fruit. I manage to get a few jars colonized completely but the mycelium was so weak that once shaken to see the recovery rate after fully colonized, it just completely stalls or recovers so slowly that putting it in CVG seems to do nothing. The seeds completely stop recovering and start producing metabolites and even so much water that you can see rings of water around the seeds after the shake once they've reached full colonization.


Here is what the agar looked like that was made with the same syringes that made the jar above.


Here is a pic of a "fully colonized" tub, obviously not fully colonized yet its forming hyphal knots. Made from seeds that did exactly what I mention above.



Here are the lids that I use. The polyfill is stuffed as tight as I can manage. Some of them so tight that the needle is making squeaking noises as I place it through the polyfill during inoculation.



These were made about 3 weeks ago with a syringe I fully expected to fail and do nothing. Same batch of syringes that made the first pic. Its basically completely stopped growing at this point. Its looked like this for about a week now. They were made with WBS that had a decent amount of corn in it. If I were to shake these they would likely not take hold of new seeds around the colonized ones. Plus its no where near a time one would shake this. These are just a couple of the jars, some of these only have growth on one side of the jar. 



These are the last jars I inoculated that are 7 days old with not even the slightest growth. (I know thats average time til growth is first noticeable) They were made with Pennington Classic WBS that has next to no corn. Pennington Classic is what I use 99% of the time. I zoomed in on one of the inoculation sites on one of them, like super zoomed in. I suppose I should have cleaned the outside of the jar much better before taking the pic. The spots you are seeing are what I assume to be dried water or myc starting to germinate or starch that may be dried on the inside of the glass. Regardless, I see this even in successful jars as they dry once removed from the pressure cooker. Yes they were pressure cooked at 15 PSI for 90 minutes.

Sometimes it can take weeks so I'm not saying these wont work but usually I see at least some tiny spot of myc after 4 or 5 days post inoculation via MS syringe. These are also made with recently purchased syringes so the syringes haven't been sitting around for more than a couple days before inoculating em. I plan to keep updating this with progress as I notice it in relation to these jars.

Before anyone says I need to rinse my WBS more, look at this and say that. 



I will be doing the whole process from start to finish in this thread. I want everyone to see every step that I take in hopes that someone can find something wrong with what I am doing. I am open to and in fact I welcome criticism.

Here is what I am currently doing. I have 7 quarts worth of pennington classic wbs that was rinsed perfectly and is seen in the pic above. They are currently sitting in hot tap water thats seen in the picture. I plan to change the water a couple of times before the 24 hour mark is reached. And I will rinsed them a few times in this process as well. Once they have soaked for 24 hours I will hopefully see a difference in the size of the seeds because I literally have never noticed a difference in the size of the seeds after soaking them. Maybe I just dont look at it very well but I do try and pinch one usually once its soaked long enough. I dont usually pressure cook them until theyve soaked long enough for me to easily pinch through one of them and see it has soaked through decently. Ill take a picture and make updates once I reach that point just to show what the seeds look like before I pressure cook them and after a soak and simmer.

Mid rinse

Finished rinse


Feel free to ask about anything I may have left out so far. I hope we can figure this out! 😅


Edited by EntheoGod (06/26/20 03:02 PM)


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OfflineEntheoGod
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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785127 - 06/26/20 04:00 PM (3 years, 6 months ago)

Im feeling like after thinking about it, that the seeds that are colonizing in the 3 week old jar could just be colonizing because the liquid inside of the syringe hit that spot and gave it some hydration. While other parts of the jar could be drier than needed.

Been doing some reading and found out that you shouldnt remove the Pressure cooker lid right after you see the pressure drop. I have been doing this so I can shake the jars around and distribute the moisture a bit better like I read in one of the older teks. I think this time I will allow the pressure cooker to cool completely on the outside then I will remove the rocker for a few minutes then I will open the pressure cooker and shake the seeds around a bit before inoculation. Obviously I will wait until the jars are cooled completely as well. This is just one of the ideas I have that could be causing this. Although still unsure why there would be so much moisture appearing in the jars that reach full colonization and then are shaken.


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785182 - 06/26/20 04:33 PM (3 years, 6 months ago)

Yeah I dont think its your jars dude. Im a total noob but I would bet its your syringe. First you can go through as many threads on cultivation forum and you would find a bunch of threads about jars failing, most of them due to being nocced with syringes. Some may be due to inappropriate prep where the grain is too wet, but thats in the beginning, not after shake. Some may be due to lid being loose, or even pc time too short or poly fill too loose, just a one or two micropore layers. But most are due to syringes. That first jar is not looking healthy at all. As long as you are soaking grains/WBS, letting them drain and then pc them for 90 at 15 PSI your jars should be golden. But if you noc those golden jars with shit you will get shit.

So you obviously have agar, you obviously know how to make agar. Why arent you noccing up a bunch of plates with that syringe? Then cleaning that growth until you get some decent and clean looking plates and then noccing your jars with agar instead of a syringe. Just build a SAB and get to it.

Also one last thing I used the search function and found that jars that produce a lot of water after colonization are bacterial. But Im not really knowledgeable so wait for someone who know better than me before doing anything.


Edited by redhandmat (06/26/20 04:41 PM)


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OfflineEntheoGod
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Re: Help me find my error in WBS prep? [Re: redhandmat]
    #26785205 - 06/26/20 04:44 PM (3 years, 6 months ago)

I have a SAB, the SAB is a ghetto looking one but it works. I spray inside it with dish soap mixed with water. I spray the entire room I am working in down with lysol spray and I wait until it settles. The foil is just there to make me feel better about leaving things inside it when Im not working with it. When I am using it I fold the foil up. The weight is on top to prevent me from moving the SAB any while using it. During the most recent inoculation of agar, I even left one of the agar plates open in the room without moving it to the SAB just to see what kind of contaminants are floating around that room. Nothing has showed its ugly little face just yet. I did this to make me feel more comfortable about inoculating the agar inside of the SAB. I will never do it in open air but it makes me feel safer knowing that the agar plate I opened up to the room for a few minutes had nothing growing in it yet.



I did exactly what you just said already, am currently waiting a few days to see if anything grows on that agar. I used the syringe that was used to inoculate the seeds that are doing absolutely nothing so far. I will post more about that as I notice changes in the AGAR. :smile:

However, if what youre saying about it being the spores is true, why would they colonize partially in a wbs jar and then just stop and not want to colonize anymore. The agar pictured is spores I bought from a non sponsor and used. The myc looked so weird that I didnt feel comfortable transferring any of it to a new plate to see what happened. Also if the spores were the problem then putting the spores on agar would still only produce weak mycelium, am I right about that? And transferring already weak mycelium would only lead to even weaker mycelium, right?

Thing is I have bought from several different vendors and had the same results. This leads me to believe something has to be happening during the prep or something like that. If its spores that are just dead upon arrival then I cant imagine every single time I buy them for them all to be duds. Especially since I have got them from different sites every time. Most recently got them from a sponsor of the site.


Edited by EntheoGod (06/26/20 04:51 PM)


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Offlineredhandmat
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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785252 - 06/26/20 05:05 PM (3 years, 6 months ago)

Let me start by saying that both your grain prep and your SAB are awesome. I dont think you are doing anything wrong there mate

Quote:

EntheoGod said:
However, if what youre saying about it being the spores is true, why would they colonize partially in a wbs jar and then just stop and not want to colonize anymore.




Simple, because of contamination. myc often literally stop growing because of molds and bacterial contams.

Now about the spores, I didnt say your spores were the problem. You obviously have gotten myc to grow on jar on agar and even after you birthed jar to coir. I said the syringe is the problem, together with the spores you have contams in there. Just clean it on agar, it takes time. MS myc looks weird as shit on germination plate, especially when there is bacterial contams with it. Just keep at it, cleaning transfering, transferring, transferring, until you get a contam free and good looking plate. Doesnt have to be rhizo growth, even tomentose but nice and healthy is awesome. Youll have your shrooms in no time. But first you need clean myc on agar, and you dont have that mate.

For grain prep I recommend Foos tek. Its essentially just boiling water, taking it off the heat source, puting WBS in the water. Let it soak for a little time. At least say 30 minutes ot an hour if I remember correctly. Maybe as little as 20 minutes is enough. Strain the water away, let WBS sit for a little while in strainer. Put in the jars. PC at 15 PSI for 90 minutes (personally I do 120-150 minutes to be safe but my pc doesnt reach 15 psi). Let the jars cool to room temperature and then drop the CLEAN agar in the jar. Thats it.


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OfflineEntheoGod
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Re: Help me find my error in WBS prep? [Re: redhandmat]
    #26785282 - 06/26/20 05:15 PM (3 years, 6 months ago)

Thanks for being so quick to respond. I will take what you said and put it to use. If the new agar ends up colonizing and showing promise I will clean it up as soon as I can to get healthy Myc. I understand what you mean about having contams present with the spores that were working. I'm just so confused about why the new syringes just recently purchased are literally doing nothing. I am waiting to see what they do on agar. I currently have 6 that were inoculated with the new syringes that I bought (golden teacher strain) I know strain doesnt typically matter just thought I would mention that as well.

I actually used a tek similar to that but you just leave the water in the bucket for 60 minutes.  Let me show you the experiment someone did and proved that in their environment the 60 minute hot water soak did better than other soaks.

https://www.shroomery.org/forums/showflat.php/Number/25012403#25012403

I did what I thought was best from this and I also had another batch I did along side this one that I soaked for 20 hours and didnt simmer. Both are showing absolutely no signs of growth yet but its only been about 7 days. Still though as I said earlier I usually see growth in about 3 or 4 days from the time of inoculation.

Thanks again so much to anyone who takes the time to give me some advice or anecdotal evidence. I appreciate it a ton! 😊


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785298 - 06/26/20 05:22 PM (3 years, 6 months ago)

Yeah I see you have poured a lot of heart in this my dude. Do not give up. I think you got this, put the same effort you did with the grain teks on the cleaning and transferring agar tek and you will make it. I cannot answer for why the new syringes are not showing any growth, but it i not unheard of spore germination to take more than 10 days! From my side I know that clean syringes simply do not exist. Using syringe to BRF usually works out well, but to grain... doubtful.

Good luck to you! and I hope more experienced people will answer your thread.


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OfflineEntheoGod
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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785301 - 06/26/20 05:25 PM (3 years, 6 months ago)

I will be sure to post pics of the seeds once they are hydrated to what I believe to be the optimal hydration. Hoping to receive some criticism there. The grains are usually translucent right up until the center of the seed once I split them with my fingernail. I try to wait until the seeds are translucent completely on the insides before deciding to load em into jars.

Thank you redhandmat, I will put as much effort as I need to into this. I thoroughly enjoy it as it gives my anxiety ridden brain something to focus on but as one can see that anxiety causes me to be a bit of a stickler when it comes to finding out exactly where this is going wrong.

I believe it is either spore related (which should be resolved once I manage to get some clean agar) or its WBS hydration related (which I will be soaking for 24 hours then doing a simmer if what I have after soaking isnt hydrated thoroughly enough)!


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785361 - 06/26/20 05:56 PM (3 years, 6 months ago)

Here is the most recent agar that I prepped and how I prepped it and what I used. If anything is wrong tell me. :smile:
Micropore tape, regular agar, Karo, and potato flakes.




I followed this tek for the agar https://www.shroomery.org/forums/showflat.php/Number/19208976

But still please break down my WBS prep to help me find what I am doing wrong. I have had tons of success with WBS and MS syringes in the past but cant get a single one to work anymore. Hoping to see something on agar as soon as possible. Its been 2 days so far after I inoculated that agar seen in the pic. I definitely know it takes longer than 2 days.


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Re: Help me find my error in WBS prep? [Re: redhandmat]
    #26785436 - 06/26/20 06:32 PM (3 years, 6 months ago)

Quote:

redhandmat said:
Yeah I see you have poured a lot of heart in this my dude. Do not give up. I think you got this, put the same effort you did with the grain teks on the cleaning and transferring agar tek and you will make it. I cannot answer for why the new syringes are not showing any growth, but it i not unheard of spore germination to take more than 10 days! From my side I know that clean syringes simply do not exist. Using syringe to BRF usually works out well, but to grain... doubtful.

Good luck to you! and I hope more experienced people will answer your thread.




Clean syringes do exist...you just shouldn't count on them...and there's no way to tell if it's clean except putting it to agar (preferred) or rolling the dice and noccing up jars and trying to spawn and fruit them.

I grew for many years with syringes > grain...then my luck ran out...and I had to learn agar. I've had literally dozens of clean syringes... until I didn't anymore.

I recently got a clean syringe from a very well known vendor... nocced up a bunch of plates and they were all clean as a whistle...I still transferred from the germ plates to be safe.

But for all intents and purposes...just assume every syringe is dirty and put it to agar first.

OP, it is most likely your syringes like the other poster mentioned. There's many...many ways to prep grain and they all work. I understand your frustration with having prior success with syringe > grain but now not being able to produce... I've been there ...trust me.

Your grain prep sounds fine... you could cook it longer..I now cook my grains 2.5 hrs @ 15psi..but again, I really don't think that's your problem.

It's perfectly normal for cube mycelium to germinate then stop growing once the contams grab a hold in the jars.

Changing your grain prep technique is not necessarily going to solve your problem. I prep my grains different all the time. Depending on how much time I have or what I'm doing. Sometimes I soak over night, drain, load and cook. Sometimes I boil for 45 minutes, sometimes I throw them in boiling water and turn the heat of for 45 minutes, somtimes I soak them over nigth then bring them to a boil. How I prep my jars for master jars, g2g receiving jars, and jars getting liquid culture are all prepped different and they all work.

It's not how you prep your grains...it's how clean the inoculant is that's going into those jars. That's going to be your number one factor leading to success.


--------------------
"Not what could have been, or what should have been, but what will be" -Self
:superbanana::mjdance::dancer:


Edited by lateforthafuture (06/26/20 06:44 PM)


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InvisibleFriedEggS
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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785437 - 06/26/20 06:33 PM (3 years, 6 months ago)

Quote:

EntheoGod said:
Here is what the agar looked like that was made with the same syringes that made the jar above.





that is your problem. i would never put that agar to grain. it doesn't look clean at all. do 4-5 transfers to new agar plates until you have a really nice looking rhizo culture. it doesn't sound like you're doing any transfers at all, are you? squirting spores onto agar and then adding that agar to grain doesn't accomplish anything unless you are taking sectors out of it to clean up the culture first.


everything else you're doing sounds fine but here is how i like to prep my WBS:
soak or no soak doesn't matter. soaking makings the simmer process quicker so i add a soak the night before. the important thing is that i simmer my WBS until the center is fully cooked. i bite a few kernels to make sure. the center should not be white and hard. simmering it will cause some grains to explode but that's normal with WBS. after it is simmered, pour out the grain into a strainer and rise it with very hot water for 10-15 seconds to remove the starch (that will help shaking your jars later). shake your strainer to remove the excess water and then let it sit in the strainer to dry in the open air for about 1 hour. then load into jars and PC.


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Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26785458 - 06/26/20 06:41 PM (3 years, 6 months ago)

Quote:

FriedEgg said:
Quote:

EntheoGod said:
Here is what the agar looked like that was made with the same syringes that made the jar above.





that is your problem. i would never put that agar to grain. it doesn't look clean at all. do 4-5 transfers to new agar plates until you have a really nice looking rhizo culture. it doesn't sound like you're doing any transfers at all, are you? squirting spores onto agar and then adding that agar to grain doesn't accomplish anything unless you are taking sectors out of it to clean up the culture first.


everything else you're doing sounds fine but here is how i like to prep my WBS:
soak or no soak doesn't matter. soaking makings the simmer process quicker so i add a soak the night before. the important thing is that i simmer my WBS until the center is fully cooked. i bite a few kernels to make sure. the center should not be white and hard. simmering it will cause some grains to explode but that's normal with WBS. after it is simmered, pour out the grain into a strainer and rise it with very hot water for 10-15 seconds to remove the starch (that will help shaking your jars later). shake your strainer to remove the excess water and then let it sit in the strainer to dry in the open air for about 1 hour. then load into jars and PC.





I didnt use that agar at all. I seen it looked like what is in the pic and I tossed it instead of even making transfers. I fully intend on having clean agar before i use any wedges to make inoculations with. I think there may have been a misunderstanding if thats what you got out of my description. I was just saying that the agar that was pictured and shows growth wasnt to my liking and i didnt trust myself to make a clean transfer with something that looked like that and it (the spores from the syringe) was performing terribly on grain as is (barely growing) so I thought that by transferring that to another plate would have only made it weaker? Am I wrong in thinking that?

All the grain jars you are seeing were inoculated using a MS syringe that wasn't on an agar wedge.


Edited by EntheoGod (06/26/20 06:44 PM)


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Offlinelateforthafuture
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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785463 - 06/26/20 06:47 PM (3 years, 6 months ago)

Quote:

EntheoGod said:
Quote:

FriedEgg said:
Quote:

EntheoGod said:
Here is what the agar looked like that was made with the same syringes that made the jar above.





that is your problem. i would never put that agar to grain. it doesn't look clean at all. do 4-5 transfers to new agar plates until you have a really nice looking rhizo culture. it doesn't sound like you're doing any transfers at all, are you? squirting spores onto agar and then adding that agar to grain doesn't accomplish anything unless you are taking sectors out of it to clean up the culture first.


everything else you're doing sounds fine but here is how i like to prep my WBS:
soak or no soak doesn't matter. soaking makings the simmer process quicker so i add a soak the night before. the important thing is that i simmer my WBS until the center is fully cooked. i bite a few kernels to make sure. the center should not be white and hard. simmering it will cause some grains to explode but that's normal with WBS. after it is simmered, pour out the grain into a strainer and rise it with very hot water for 10-15 seconds to remove the starch (that will help shaking your jars later). shake your strainer to remove the excess water and then let it sit in the strainer to dry in the open air for about 1 hour. then load into jars and PC.





I didnt use that agar at all. I seen it looked like what is in the pic and I tossed it instead of even making transfers. I fully intend on having clean agar before i use any wedges to make inoculations with. I think there may have been a misunderstanding if thats what you got out of my description. I was just saying that the agar that was pictured and shows growth wasnt to my liking and i didnt trust myself to make a clean transfer with something that looked like that and it (the spores from the syringe) was performing terribly on grain as is (barely growing) so I thought that by transferring that to another plate would have only made it weaker? Am I wrong in thinking that?

All the grain jars you are seeing were inoculated using a MS syringe that wasn't on an agar wedge.




Transferring doesn't "weaken" the mycelium...if anything it will help it by getting it away from the contams. It may be appearing weak if it's fighting against contams in your dish or jar...isolate it away from the contams and it should thrive...if that makes sense.


--------------------
"Not what could have been, or what should have been, but what will be" -Self
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Edited by lateforthafuture (06/26/20 06:49 PM)


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InvisibleFriedEggS
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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785468 - 06/26/20 06:50 PM (3 years, 6 months ago)

ok yea i misunderstood. that plate looks normal for a germ plate. taking a transfer from it won't make it worse, it'll make it better. on the germ plate it's sometimes hard to choose where to transfer from but just do your best. it may not seem like it will work but you'll be surprised. the germ plate looks blobby because there are tons of cultures growing on top of each other so you can't see any rhizo growth yet. cultures also look weird when there is bacteria in the mix. but take a few transfers and it'll slowly look better each time. take transfers from different locations and you'll increase your chances.


yea try to move away from injecting spores into grain jars and you'll have much better luck :thumbup:


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OfflineEntheoGod
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Re: Help me find my error in WBS prep? [Re: lateforthafuture]
    #26785486 - 06/26/20 06:57 PM (3 years, 6 months ago)

Thank you. I agree and am working diligently to produce some clean agar. So in your opinion do you think even though the jars that I inoculated with the new syringes aren't doing anything, would the spores give me some results on the agar I already have them on? Or do you think I could have potentially got a shitty syringe that was completely dead? I highly doubt id get a completely dead syringe so I would assume through putting them on agar I could get some results. The agar that I prepped and have inoculated is pictured in a few posts above this one.

I will still be attempting this last syringe I have on grain but I wont b using the whole syringe so I will likely use whats left to put some more on agar and clean up even more plates. I have the patience I was just hoping to have something coming in considering I havent in months. Its kinda demotivating like I said but I will not be getting too down and giving up. I do believe I will work on getting agar going before I try anything else though. After this batch I have soaking currently that is. Only because I dont wanna waste the seeds I put effort into today. The plan is like I said to do the 24 hour soak and simmer for a few minutes or until I notice the seeds arent dry inside then using a fresh syringe to inoculate when ready. I wont be using the whole syringe so I will make even more agar with whats left. Seems like the smart thing to do.

Also just noticed you replied before I could get this posted so ill respond here to both of you friedegg and lateforthafuture. Its great to know that the mycelium wont weaken through transferring it off of a germ plate. Sad that i tossed those last ones now that I hear all this :frown: Oh well I must move forward and hope I get better results. Also in that germ plate you seen, i accidentally used much more spores per plate than i should have. I accidentally shot pretty much 1/4 cc in each of those. I made sure to put as little as I could on the newer plates! 💖 Any other input is valued greatly folks. I feel like I have learned so much from your posts already.

I thought that the mycelium being weak in the WBS jars would have showed me that if it was weak on agar then there would have been no point in transferring to clean it up. I feel dumb for tossing it now. :facepalm3:


Edited by EntheoGod (06/26/20 06:59 PM)


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785508 - 06/26/20 07:05 PM (3 years, 6 months ago)

One tip next time you nocc up some plates.

Either order some sterile swabs or sterilize some q tips. To sterilize the Q tips just put them in a jar with a lid and run them in your pressure cooker.

When you go to nocc up your plates...squirt a little bit of solution onto the Q tip ...then streak the plate with the Q tip. I've had really...really good success with this technique. Or you could use an inoculation loop. Streaking the solution will help you see exactly where you inoculated when the growth starts..it will be easier to tell what's mycelium and what's a possible contam...at least in my experience. It will also reduce the amount of spores you get onto your plates..it's really hard to control how much you put on the plates just squirting a syringe on them.


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785511 - 06/26/20 07:07 PM (3 years, 6 months ago)

Actually thinking about taking one of those colonized seeds from the shitty jar and getting it to germ on a new plate once I get more supplies in. Wonder if that could help me try cleaning it up or would you guys rather just start with ms syringe instead? 🤔

I have some sterile swabs in my amazon cart right now, ordering it asap. its the 100 pack https://smile.amazon.com/gp/product/B003IT74FO/ref=ox_sc_act_title_1?smid=A29OWEYSFJVSZC&psc=1

I think I will try sterilizing my own swabs as well just for convenience sake.

I never thought to put the spores from the syringe on a swab. I always thought that was only for putting prints to agar! I am learning so much. I guess it makes sense considering I just recently started agar due to all of these failures.


Edited by EntheoGod (06/26/20 07:13 PM)


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785532 - 06/26/20 07:15 PM (3 years, 6 months ago)

i think your syringe just has some bacteria in it. maybe it came that way or maybe bacteria got in there from reusing the syringe improperly several times.

your agar prep looks fine :thumbup:

grains are not very tolerable of bacteria. yes too much spore solution is a bad thing. you only need 2 drops on a plate. if you really want to get something going with your syringe right now, try the PF tek while you wait for clean agar growth. cakes can tolerate bacteria better than grain. it won't be a lot faster but it's worth a try.



if you want some advice to make your syringe stay cleaner and last longer, here you go:
your spore syringe should have came with a needle and a syringe tip cap.
here is what a syringe tip cap looks like:


don't use the needle at all. when you want to use the syringe, wipe the exterior with alcohol, then remove the tip cap, carefully squirt 1-2 drops into your agar plate, and quickly put the tip cap back on. a lot of people will use the needle for some reason but flaming the needle and squirting solution out of the syringe to cool the needle is a waste. the needle is also a vector for contamination if you leave it on during storage. if you only use the tip cap and only use 1-2 drops per plate, it'll last a very long time.


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Edited by FriedEgg (06/26/20 07:19 PM)


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InvisibleFriedEggS
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Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26785537 - 06/26/20 07:17 PM (3 years, 6 months ago)

one more thing. since you are in a hurry to get clean growth, you can sprinkle some of those colonized grains onto a new agar plate. they will rapidly grow out and you should have a clean culture in only a week or two. :thumbup: when spores are stubborn to germinate on agar that's how you get clean growth anyways.


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Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26785547 - 06/26/20 07:20 PM (3 years, 6 months ago)

Quote:

FriedEgg said:
one more thing. since you are in a hurry to get clean growth, you can sprinkle some of those colonized grains onto a new agar plate. they will rapidly grow out and you should have a clean culture in only a week or two. :thumbup: when spores are stubborn to germinate on agar that's how you get clean growth anyways.




Agreed...I'd pluck some of those grains and drop them straight to agar...it shouldn't take long at all to clean that up.


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785554 - 06/26/20 07:24 PM (3 years, 6 months ago)

Quote:

EntheoGod said:
Here are the lids that I use. The polyfill is stuffed as tight as I can manage. Some of them so tight that the needle is making squeaking noises as I place it through the polyfill during inoculation.





one more thing


those jars will work fine but they aren't necessary. i use unmodified plastic lids. to use them, screw the lid on tight, and then unscrew slightly about 1/8th of a turn. it should be snug but loose enough to allow a little gas exchange. if the lid is too tight your growth will stall at about 50% but can be reignited by unscrewing the lid more. if it's too loose it'll either dry out or get contaminated. at first it sounds like it's a recipe for contamination but trust me, it's not.
set the jar somewhere and don't disturb it. be careful when picking it up. don't handle the jar by the lid.



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Edited by FriedEgg (06/26/20 07:31 PM)


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Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26785572 - 06/26/20 07:31 PM (3 years, 6 months ago)

I will wait until I get clean agar, I dont think BRF would be much faster than me simply working with the agar and getting it to grain once its cleaned up. I flame the needle after use and before putting the cap back on it. But I havent re used a syringe several times ever. I only recently used it to inoculate a set of agar plates because I had some spores leftover and didnt have the syringe cap to replace the needle after I used it. From what I understand, storing the leftover syringe would be bad but it would b much worse if i were using that leftover syringe on the grains, which Im not. If theres any bacteria in the syringe due to the needle and me storing it for a couple of days then I could easily get it clean through agar. This is all assuming I am applying the tips that everyone has given me already on here. I mean if I were to believe everything I have read then I should  also assume that even bacterial syringes can produce healthy spawn when put to wbs. NOT GOING TO BE DOING THAT but its been said on here by TC's along with long time growers who arent TCs.

I store the leftover spores inside the original package they come in. They come in a plastic bag thats inside of a box. After using the syringe with the needle. I flame the needle and while its relatively hot i then place the cap for the needle back on and put it immediately inside the plastic bag and place that inside my drawer thats in the same room as everything else. I was told that would be safe as long as I wasnt using it on grains after storing that way. And I am definitely not doing that.  BTW the way I flame the needle isnt a simple lighter. I use a butane torch. I get that when I say I flame the needle it sounds like Im just putting it to a flame and hoping that kills anything but I torch it til red hot.




I look forward to seeing something clean and beautiful come from all this work. I want to see a beautiful agar plate with some nice rhizomorphic growth.

EDIT: Again I noticed you guys posted before I could type out this book of a post lol. I will do that as soon as my new plates come in and while im waiting for the ones I have currently to show me any signs of growth. I look forward to this so much right now. Im getting excited just thinking about all the possibilities :smile: Ill keep to the basics since I am a beginner though. As soon as those new plates come in the mail ill be doing exactly what you said and dropping a colonized grain on a few of them and cleaning that up from there. Once I have something clean the point will be to prep some grains like I normally would and noc them up using a wedge or a few wedges per jar.

The lids you mentioned do look nice. Especially if I completely stop inoculating via syringe but I do like the not having to open the entire lid to inoculate. I was hoping to get good enough that I make Liquid inoculant eventually and that would feel much safer to me, if I were to inoculate it through the lid like I have already. Now that I am thinking about it, it sounds like im just making excuses to not get those lids. Ill get em for sure. If I have to modify them at all then so be it. Not seeing why I couldnt just open the lid inside my SAB to do the inoculating via Agar wedge or LI.


Edited by EntheoGod (06/26/20 07:37 PM)


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785611 - 06/26/20 07:44 PM (3 years, 6 months ago)

storing a leftover syringe is fine as long as you don't contaminate it. that's why i explained to use the tip cap and not the needle. i think storing it with the needle still attached is what caused all my problems in the beginning. i have syringes that are a year old and they are probably fine.


yes, even bacterial syringes can produce healthy spawn when put to wbs.... but it's not reliable or consistent :wink:


As soon as what new plates come in the mail? i thought you already have plates

instead of LC, look into G2G (grain to grain). LC is hard to work with and can cause bacteria problems if not done properly. G2G is much more reliable.

those polyfil lids are really only used for injecting into grain jars. to do A2G (agar to grain) or G2G, you'll need to open the lids anyways. and don't worry, opening a lid isn't that risky in a still air box.

good job so far. you'll be there soon :thumbup:


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Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26785619 - 06/26/20 07:48 PM (3 years, 6 months ago)

Yea I already have like 7 plates but theyre a bit occupied at the moment with agar that I have inoculated with a newer syringe and am waiting for them to germinate. I have more plates coming in the mail and when i get those Ill prep up some more agar and drop some grains onto it if nothing else turns out better for me in the meantime.


Gotcha about the LI. Ill hold off on that until I get way better and more consistent results. I just see the growth rates of LI and it makes me so jealous. HAHA. I cant help it. But for the better of the seeds Ill stick to developing my agar and cleaning it up. Doing a few transfers and then inoculating via agar wedges. Once the jars I inoculate with agar wedges do well I will then try doing G2G. This feels like itll help a lot with cleaning up my act so to speak. 🤣


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785629 - 06/26/20 07:53 PM (3 years, 6 months ago)

you're on the right track now :thumbup:


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Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26785636 - 06/26/20 07:57 PM (3 years, 6 months ago)

btw i just seen that amateur mycologist united link in your sig. I appreciate people like that. Thank you. I may end up using that sometime soon if I have much trouble with anything else. I will continue to update this thread with the progress as it happens.

Next post on this thread will be when I simmer the seeds I have soaking. I am about to go change out the water its been soaking in for about 9 hours for hot water again. Also gonna toss a bit of gypsum in the soak. Not much just a pinch since I am about completely out. Gotta get more soon or might not since I have seen its not needed at all. Gonna post about the seeds and their hydration in the next update to this thread. I will show a seed before soaking and after to show what happens to my seeds in 24 hours. I have read they are suppose to double in size about. I have never, in the 5 years ive been doing this, ever seen my seeds double in size after hydration. It always looks about the same to me. Im hoping to change that with this batch. :smile:


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785664 - 06/26/20 08:12 PM (3 years, 6 months ago)

G2G can be as fast or faster than LC. If you only put 1-2cc of liquid culture it's probably going to take a few weeks to colonize. If you put 5cc + of LC it will be much faster.

I love G2G, it is crazy fast... you'll have more clean spawn than you know what do with. If I G2G 1 qt to 10 qts...they are usually done in 7-10 days. If I do one qt to 7 or even 5 qts...they are done in under a week.

LC vs G2G speed is more about how much inoculate you use, IMHO. If you want your jars done fast... use more inoculant (G2G or LC).

I like G2G because I can visually see what's going into my receiving jars...kinda hard to tell with LC, especially if it's been sitting a while. I have some oyster liquid culture that looks crazy...but I just nocced up some jars with it and it's fine.


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Re: Help me find my error in WBS prep? [Re: lateforthafuture]
    #26785691 - 06/26/20 08:30 PM (3 years, 6 months ago)

I am just stoked at the thought of not encountering contaminations as often or ever. As well as being able to know that what I put in the grains isnt bad. I do have this question though while we're here. How long, once started germinating on the agar, should I wait until transferring to another new plate? I was thinking you should let the mycelium grow for a few days to a week before trying to transfer anything to another plate but is that wrong? I havent taken the time to search this yet so I should probably just do that.

I have all the time in the world to do this so whatever happens happens but I will do everything in my power to prevent anything bad. I am just hoping I can pull this off with as little contams as possible in the agar. So far so good. Even on the first batch of agar that I tossed due to me being an idiot, I didnt see any bad contaminations. 👍 You live and you learn. I love that about this hobby. I've learned a ton today alone, so much so that I am still trying to get it all in my memory. I have a notepad with tons of quotes from TC's about agar and even a few from this thread alone with you guys helping me!


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Re: Help me find my error in WBS prep? [Re: lateforthafuture]
    #26785697 - 06/26/20 08:32 PM (3 years, 6 months ago)

@Friedegg (or anyone) Is the simmer after soaking necessary? Wouldn't the grain be more or less cooking in a PC, or while steaming? Seems like an unnecessary step.


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785710 - 06/26/20 08:41 PM (3 years, 6 months ago)

Quote:

EntheoGod said:
I am just stoked at the thought of not encountering contaminations as often or ever. As well as being able to know that what I put in the grains isnt bad. I do have this question though while we're here. How long, once started germinating on the agar, should I wait until transferring to another new plate? I was thinking you should let the mycelium grow for a few days to a week before trying to transfer anything to another plate but is that wrong? I havent taken the time to search this yet so I should probably just do that.

I have all the time in the world to do this so whatever happens happens but I will do everything in my power to prevent anything bad. I am just hoping I can pull this off with as little contams as possible in the agar. So far so good. Even on the first batch of agar that I tossed due to me being an idiot, I didnt see any bad contaminations. 👍 You live and you learn. I love that about this hobby. I've learned a ton today alone, so much so that I am still trying to get it all in my memory. I have a notepad with tons of quotes from TC's about agar and even a few from this thread alone with you guys helping me!




I would transfer as soon as you see enough healthy mycelium that you can grab.  The longer you wait to transfer the more chance a contam has to present itself. Get the healthy mycelium to a new clean plate ASAP. I couldn't give you an exact time frame but it's days not weeks.


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Re: Help me find my error in WBS prep? [Re: checkmatsis]
    #26785718 - 06/26/20 08:44 PM (3 years, 6 months ago)

Quote:

checkmatsis said:
@Friedegg (or anyone) Is the simmer after soaking necessary? Wouldn't the grain be more or less cooking in a PC, or while steaming? Seems like an unnecessary step.





It isnt necessary. The only thing thats necessary is getting your grains properly hydrated. :smile: Thats what I gather from all the information I have sifted through over the past few days. Some soak for 24 hours and no simmer, some soak for 10 hours, some only simmer and dont soak.

Its all about proper hydration of your seeds and obviously clean spawn.

I have read though, that it does help in certain situations, like if your seeds arent getting fully hydrated before tons of endospores germinate in the soak. Soaking past a certain point is detrimental to the seeds. You soak until the seed is either fully hydrated and you dont need to simmer, OR you soak until you reach a point that you are possible germinating too many endospores for the pressure cooker to kill off. Which happens, from what I have read, around 36 hours for most people.  I believe thats why its so promoted on here that you soak for 24 hours then simmer. If your seeds are fully hydrated in 24 hours then a simmer wont help much other than drying the outside of the seeds.


Edited by EntheoGod (06/26/20 08:48 PM)


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Re: Help me find my error in WBS prep? [Re: lateforthafuture]
    #26785722 - 06/26/20 08:45 PM (3 years, 6 months ago)

Quote:

lateforthafuture said:
Quote:

EntheoGod said:
I am just stoked at the thought of not encountering contaminations as often or ever. As well as being able to know that what I put in the grains isnt bad. I do have this question though while we're here. How long, once started germinating on the agar, should I wait until transferring to another new plate? I was thinking you should let the mycelium grow for a few days to a week before trying to transfer anything to another plate but is that wrong? I havent taken the time to search this yet so I should probably just do that.

I have all the time in the world to do this so whatever happens happens but I will do everything in my power to prevent anything bad. I am just hoping I can pull this off with as little contams as possible in the agar. So far so good. Even on the first batch of agar that I tossed due to me being an idiot, I didnt see any bad contaminations. �� You live and you learn. I love that about this hobby. I've learned a ton today alone, so much so that I am still trying to get it all in my memory. I have a notepad with tons of quotes from TC's about agar and even a few from this thread alone with you guys helping me!




I would transfer as soon as you see enough healthy mycelium that you can grab.  The longer you wait to transfer the more chance a contam has to present itself. Get the healthy mycelium to a new clean plate ASAP. I couldn't give you an exact time frame but it's days not weeks.





Sounds like a plan. Ill be sure to do that as soon as I see myc that looks good enough to grab. Im sure ill post on here to gather opinions from folks like you beforehand. I like getting opinions first especially since I am new to agar.


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785817 - 06/26/20 09:36 PM (3 years, 6 months ago)

Update: The 7 qts of wbs that I have soaking have been soaking for around 10-11 hours, I decided to change the water out for new hot water.



Before rinsing a few more times and changing the water.


After


I didnt throw in any gypsum because well, I am about completely out and decided to save what I have for the CVG when I need it. Now to wait until tomorrow around 12 noon and then start a short simmer (if needed).

Here is what the grain looked like on the inside after the short 11 hour soak so far. Couldnt get the cam to focus but you can still see what I mean.


I want the grain to be translucent all the way through, you can see its getting towards the center but isnt soaked through just yet. Another 11 -12 hours should do it but maybe not for all the grains so I am likely still going to simmer when the soak is up.


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Re: Help me find my error in WBS prep? [Re: checkmatsis]
    #26785897 - 06/26/20 10:12 PM (3 years, 6 months ago)

Quote:

checkmatsis said:
@Friedegg (or anyone) Is the simmer after soaking necessary? Wouldn't the grain be more or less cooking in a PC, or while steaming? Seems like an unnecessary step.




some people have success without simmering but i dont. i don't like doing extra work so trust me i've tried avoiding the simmer step. but whenever i do, the top 20% of my grain won't colonize because it dries out. i've tried soaking 24 hours and still adding some water to my jars but all that does is make the bottom of the jar mushy and the top of the jar underhydrated.


this is the shroomery member "gr0wer" and he doesn't simmer. but he only grows edibles. oysters might be able to tolerate it more than cubes. i dont know.


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Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26786376 - 06/27/20 05:21 AM (3 years, 6 months ago)

@EntheoGod Yeah lots of variety in grain preparation and all seem to yield results as long as (as you said) hydration and sterilization are properly achieved. I'll be soaking for 12 hours, simmer (still out on this), 45-minute drain, then steam sterilize for 6 hours. This should allow for proper hydration and sterilization in my experience.

@Friedegg No extra work for me either lol. Wouldn't shaking your jar after some colonization is achieved help mitigate the dry top and moist bottom and allow for a more full colonization of your WBS jars? Thanks for the video.


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Re: Help me find my error in WBS prep? [Re: checkmatsis]
    #26787270 - 06/27/20 12:15 PM (3 years, 6 months ago)

Quote:

checkmatsis said:
@Friedegg No extra work for me either lol. Wouldn't shaking your jar after some colonization is achieved help mitigate the dry top and moist bottom and allow for a more full colonization of your WBS jars? Thanks for the video.



it didn't help when i tried


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Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26787411 - 06/27/20 01:24 PM (3 years, 6 months ago)

I can only imagine how many people have read this post and decided not to reply.

Op: are you venting your PC and then letting it cool overnight before removing the jars?

The agar: do you have a picture of the clean agar you are planning on using next.

The syringes: unless your getting them from a vendor with a centerfugue, no syringe is clean.

Your getting bacteria in your spawn. So there are many ways of that happening. Not getting a proper PC run, usually an air bubble in the Pac preventing the steam from fully sterilizing your jars. Could also be faulty lids. Could be bad inoculate.

Just skip syringes to grain cause that will always be the biggest failure point. It has worked before for people, but if your trying to get clean and healthy spawn. Agar to properly sterilized grain is the best way to go.

Also are you using a flowhood or SAB?

Grain prep: load up a pot 3/4 full of water and bring it to a full rolling boil. Then add WBS no rinsing just dump it in.

After about 20 minuets kill the heat and strain immediately. Then lay out on a beach towel or news print in a large even surface to steam dry. Give it an hour and don't forget to stir the grains to allow all of the steam to dry the outside of the grain. Then load and PC.


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Re: Help me find my error in WBS prep? [Re: Sockadin]
    #26787568 - 06/27/20 02:21 PM (3 years, 6 months ago)

Quote:

Sockadin said:
I can only imagine how many people have read this post and decided not to reply.

Op: are you venting your PC and then letting it cool overnight before removing the jars?

The agar: do you have a picture of the clean agar you are planning on using next.

The syringes: unless your getting them from a vendor with a centerfugue, no syringe is clean.

Your getting bacteria in your spawn. So there are many ways of that happening. Not getting a proper PC run, usually an air bubble in the Pac preventing the steam from fully sterilizing your jars. Could also be faulty lids. Could be bad inoculate.

Just skip syringes to grain cause that will always be the biggest failure point. It has worked before for people, but if your trying to get clean and healthy spawn. Agar to properly sterilized grain is the best way to go.

Also are you using a flowhood or SAB?

Grain prep: load up a pot 3/4 full of water and bring it to a full rolling boil. Then add WBS no rinsing just dump it in.

After about 20 minuets kill the heat and strain immediately. Then lay out on a beach towel or news print in a large even surface to steam dry. Give it an hour and don't forget to stir the grains to allow all of the steam to dry the outside of the grain. Then load and PC.





Why would you say people are reading the post and deciding not to respond?

About the Venting, no I have not been doing that properly as I stated in a few posts previously. I have read about this just a couple days ago and thats one of the biggest reasons I have WBS being pressure cooked right now. I plan to do this with the batch I have currently being pressure cooked. I will remove it after around 90 minutes, might let it go for longer today, then I will place it in the room where everything else is kept. I allow it to cool in that room until the pressure drops just enough for the pressure gauge to go to 0 and the air vent/cover lock, as well as the overpressure plug to drop. Then I will usually immediately take the jars out of the pressure cooker while still hot to the touch, shake them around to distribute water thats built up and I place them immediately inside of the SAB until time to inoculate them. I allow them to cool completely inside the SAB. I know now that this is probably the wrong way of doing it so this time with the batch I have going I will do as you said, and let the pressure drop completely then remove the pressure regulator to allow it to vent properly and allow everything to cool inside the cooker overnight until its completely cool jars and all.

When I am pressure cooking the seeds, I always make sure to let steam vent from it for a few minutes before placing the weight on top. I wait until I see pure steam pushing out then place the weight on and it goes to 15psi and I turn the heat down to medium and let it sit there for 90 minutes or more, like I said.

About the Agar, I have pics of the ones that were most recently inoculated that I didnt think were bad. I decided to toss a whole batch that evidently could have been used to transfer and clean up the myc. Its only been a few days since I inoculated this agar in this picture. 



You cant really see the agar but I dont feel safe opening the lids to take a picture of it. I can show you a pic of what the agar I previously made which looks identical before inoculation.


I did accidentally use about 1/4 cc per agar plate which I know is too much, the syringe was pretty tough to push down and everytime id try getting a single drop ready, it ended up pushing out much more than I needed. Even ended up squirting some of it on the bottom of the SAB by accident but still managed to inoculate the plates with much less than my first attempt. I am still waiting to see results but its only been a few days so far. Like 2 days or 3.

I am using a ghetto looking flowhood tbh. But it gets the job done. I left one agar plate open to the air for a few minutes and closed it to see what contaminations may have fell inside it but nothing so far. The reason I did this was because It made me feel better even using the SAB I have. This is the SAB. The aluminium flaps are just there to make me feel better about leaving things inside it when I am not around. I dont have the means to make a proper flow hood at the moment. Money has been super tight.


I make sure to spray soapy water inside before placing the things that are sterile in it. I follow proper SAB sterile technique to a T with the SAB I have. I even use it with the Grain jars when I inoculate those. The grains that I showed earlier that had no signs at all after about a week are finally starting to show the slightest signs of growth. Nothing to get excited about though. Ill post a pic of it.(I cant get this photo properly oriented -__-)


About the Syringes, I am not an idiot so I doubt the vendors I am purchasing from have a centrifuge. I could be wrong. Not that likely though.

I have tried that grain prep as you mentioned above and it did next to nothing to hydrate my seeds through. Hell a 24 hour soak doesnt even hydrate them fully. Which is why I decided to start simmering again.

I noticed the seeds were rather dry on the inside but when I simmer them I go from this.



To this after a simmer.


Noticing barely any burst seeds.

After simmering this latest batch I placed them on a towel to dry until dry enough to pass the TP test.


They fall right off the TP without sticking at all. I noticed they werent nearly as sticky this time as well. Likely due to me changing the soak water several times and rinsing the seeds several times throughout the soak. The simmer seems to have made a difference there too. I made a batch that wasnt simmered and noticed that it was rather sticky before simmering and lost that quality afterwards.


I am absolutely going to be working for clean spawn now, after this batch of seeds I have in the pressure cooker right now is inoculated and waiting. I wont be making any more until I am able to get the agar cleaned up properly.

I follow these teks when it comes to making the agar and pressure cook it for roughly 40 minutes, following the same steps I do for grain when it comes to waiting for the pressure to drop inside except I paid more attention to the agar tek and did actually properly vent the pressure cooker then.
https://www.shroomery.org/forums/showflat.php/Number/19208976
https://www.shroomery.org/forums/showflat.php/Number/23188000

I understand no syringe will be bacteria or contam free. I know syringes aren’t clean. I am only using the syringe on this batch of seeds because I can without wasting much and I can use the leftover to start more agar considering I’ll only be using 7ccs of this syringe today. I have to make this known because a lot of people keep saying don’t use the syringe. I won’t be after today. I promise that. I need to clean up some agar and start doing it that way. Don’t get that the wrong way though I absolutely love that you all have been so helpful! 😃 just have to make that known.

I’ll be showing every single step I take with what I do today once I’ve completed everything up until inoculating.

I really appreciate you stopping by to give your input and advice. :smile: Any help given is very much needed and ingrained in my brain (no pun intended)


Edited by EntheoGod (06/27/20 02:45 PM)


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OfflineSockadin
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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26787604 - 06/27/20 02:42 PM (3 years, 6 months ago)

Sounds good. The only thing I would do differently is leave the weight on the vent pipe and let the PC cool naturally over night. If you pull it off you can create a vacuum as the pressure stabilizes and pull open air into the PC. Not the worst thing in the world cause the poly would ideally catch and contamination. But just a best technique kinda deal for me.

The reason I said alot of people probably read this but didn't respond is because in my opinion early in the thread there was some iffy advise given to you. Dare I say bad.


Also there are vendors who have centerfugue but no plugging sponsors so I won't say who.


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OfflineEntheoGod
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Re: Help me find my error in WBS prep? [Re: Sockadin]
    #26787643 - 06/27/20 03:05 PM (3 years, 6 months ago)

I gotcha yea that makes sense. I try using my own sense when doing this as well so I’ll be doing exactly what you see me describe in the last post. I would think if someone sees bad advice being given they should feel obligated to correct it but that’s just how I am. I want everyone to have a good experience of this!  I wish I knew who had the centrifuge cause that’s all I’d buy from but that’s likely why we can’t plug sponsors on here and considering they pay for advertising equally. Lol  but please if anyone sees bad advice feel absolutely free to correct it and me. I love finding out facts and if I’m doing something wrong I NEED to correct it. But as for this batch today I’ll do exactly as you said and let it completely cool in the pc overnight I just worry about not shaking the jars while hot to disperse built up water. I suppose I could still easily do this once it’s cooled overnight! :smile: I also get paranoid about leaving the foil on top because it makes me fear water building up under the foil and on top of the lid. Probably not as big of a problem as I imagine.


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Offlineredhandmat
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Re: Help me find my error in WBS prep? [Re: Sockadin]
    #26788859 - 06/28/20 04:07 AM (3 years, 6 months ago)

Quote:

Sockadin said:
The reason I said alot of people probably read this but didn't respond is because in my opinion early in the thread there was some iffy advise given to you. Dare I say bad.




Hmm isnt it better to correct that instead of just saying it like that? If I have given advice that is so bad I would love to know so that I may not say the same thing again in ignorance. Likewise if its someone else. But going through the initial posts, what I see, OP was 100% sure it was his grain prep. He acknowledged that he had tried many different teks for grains all of them failed. So I pointed out that syringe is more likely to be the culprit. Obviously OP seems to follow teks very well, and sure it could be the jars themselves or the PC etc, which was also achnoledged by repliers. But usually when troubleshooting you start from the beginning and work your way forward. Or you can start from the end and work your way backward in the process. Either way you end up with the syringe as a question mark. And thus I gave OP the same suggestions trusted cultivators in these forums have given me: AGAR.

Edit: I hope I dont come across as an asshole or anything, Sockadin. You have previously given me awesome advice that helped me. And also OP is obviously struggling like many of us here, and I would not wish to make that struggle harder. My point is that I would love to be corrected if I have given bad advice, similarly if someone else gave bad advice its good for all of us to know so we may avoid it :smile:


Edited by redhandmat (06/28/20 04:45 AM)


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OfflineSockadin
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Re: Help me find my error in WBS prep? [Re: redhandmat]
    #26789055 - 06/28/20 07:18 AM (3 years, 6 months ago)

Not you bro i just disagree with changing the cap and needle over and over again.

I reread it and might have misunderstood what he was saying. I think now he was saying to just using the syringe cap and skipping the needle when dropping water on the germination plate.


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Offlineredhandmat
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Re: Help me find my error in WBS prep? [Re: Sockadin]
    #26789827 - 06/28/20 01:22 PM (3 years, 6 months ago)

Quote:

Sockadin said:
Not you bro i just disagree with changing the cap and needle over and over again.

I reread it and might have misunderstood what he was saying. I think now he was saying to just using the syringe cap and skipping the needle when dropping water on the germination plate.




Ah alright! yeah Im a noob and still learning a lot so I wasnt sure. Anyways, I hope OP gets some nice results with agar soon :smile:


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InvisibleFriedEggS
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Re: Help me find my error in WBS prep? [Re: Sockadin]
    #26790373 - 06/28/20 05:00 PM (3 years, 6 months ago)

Quote:

Sockadin said:
I reread it and might have misunderstood what he was saying. I think now he was saying to just using the syringe cap and skipping the needle when dropping water on the germination plate.



:thumbup: exactly. you don't need a needle to squirt a little drop on a plate. it's just another contamination vector if you store it with the needle still attached (which most people do because they don't have extra needles).


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OfflineEntheoGod
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Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26791059 - 06/28/20 10:38 PM (3 years, 6 months ago)

Hey guys still working hard at it gonna be updating everything soon with pics and the progress. Just to be brief I have nothing growing in the agar I had made after tossing the first batch. No contams or mycelium. And a few of the jars are showing something but I’m fairly sure those are too dry.

The latest batch of seeds is too early to say anything but I followed the advice of fried egg when it comes to keeping the jars inside the pressure cooker til morning then inoculating them. Again that is the last spore syringe to grain I’ll be doing but had to not waste what I already had going!

I’ll be making new agar as soon as the new plates come in the mail and I’ll be dropping a seed that’s already colonized, onto a fresh agar plate to clean that bacterial myc up I’ll likely try using a pin or some mushie tissue from the only monotub I have going at the moment. And also gonna use the last bits of some syringes I have but I’ll put those on agar as well to try getting something clean and if nothing else, at least I learned something :smile: any more advice would be great as well as if you think 4 or 5 days is a long time seeing nothing on agar. No contams ,just nothing and I can see the spores that were placed on it. The agar looks a bit sweaty tbh. I’ll post pics soon as I get some more progress made towards clean spawn!


Edited by EntheoGod (06/28/20 10:40 PM)


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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26794709 - 06/30/20 02:33 PM (3 years, 6 months ago)

I have a quick question and thought it best to share it on this post as opposed to making an entirely separate one.

Ive been seeing different agar pressure cooking times being thrown around and would like to get a sort of consensus or at least someone to tell me if I did it too long. I pressure cook the PastyPlates at 15psi for 45 minutes. I am noticing literally no growth in the latest agar plates I made. Not even a contamination. You can clearly see the spores yet nothing seems to have changed since day 1. Its been around about 5 or 6 days since inoculating them with as little spore solution as possible.

If I am being impatient I have no problem waiting at all. Just wanted to check and see if I should scrap those and make fresh agar so I could pressure cook for a shorter time. Not thinking that would be the problem but I thought it best to ask and get some opinions.

From my perspective, I think I am doing it properly and nothing should be wrong (other than potentially bad or dead spores) I have no idea why every spore in a syringe could be dead so I dont think that would really be a problem either.

For all I know, I have done everything right and just need to wait for results. Regardless, I will be making 8 more pasty plates today and was hoping I could still get a consensus on how long to pressure cook them. I see on his tek it says 40-45 minutes and then I see other TC's saying 20 minutes is all that is needed because any more could cook the sugars inside the agar. :eek: All these different times and different information is a bit confusing and makes me question whether I am doing it right to begin with..

I feel that I have to reiterate that I am not really being impatient so much as I am unaware of what to expect around what time frames really. I am willing to wait as long as needed. I keep reading a variety of different responses to older posts that say several different things. Some say I should wait 20 days some say if you dont see growth on agar within a week to scrap it. 

(gonna be using colonized grains or pins to inoculate the agar I plan on making soon.)

Thanks a ton for your help!


Edited by EntheoGod (06/30/20 02:38 PM)


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Offlinecheckmatsis
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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26795034 - 06/30/20 05:08 PM (3 years, 6 months ago)

Not sure if you've already seen this post, but:

https://www.shroomery.org/forums/showflat.php/Number/17052521


"Spores can take up to 3 weeks to germinate...even on agar." - PussyFart


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OfflineEntheoGod
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Re: Help me find my error in WBS prep? [Re: checkmatsis]
    #26795270 - 06/30/20 06:58 PM (3 years, 6 months ago)

Yea, I absolutely said I understand that it can take a while for anything to germinate. Where I am new to agar I trust a majority of what I read on here when its posted by TC's but the varying degrees of information can be pretty tedious to sift through. I find it difficult to understand whether its a bad thing if it takes more than a week to germinate or whether that's common for agar or spores in general when I come across multiple different answers to the same question. I do understand that everyone has different levels of success so the experience is subjective to a degree. But what would be considered the typical germination times for spore solutions put to agar? Like if you were to ask me when you should see growth on something like a spore syringe put on grains, I would, subjectively, say that if you don't see growth of some sort within 4 days then something is up. But again that's completely subjective and I know that its a fact that spores can take much longer to germinate. They CAN but is it a good sign if they do?

I hope I don't sound too high to have replied yet 🤣


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Offlinetripdawg420
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Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26795331 - 06/30/20 07:21 PM (3 years, 6 months ago)

im sorry i didnt read all this whats your problem


--------------------
HUSTLER
How U Survive This Life Everyday Resourcefully
epic GT mono tub
http://www.shroomery.org/forums/showflat.php/Number/17277772

wbs tek
http://www.shroomery.org/forums/showflat.php/Number/11525679
coir tek
http://www.shroomery.org/forums/showflat.php/Number/11917410
results :thumbup:


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OfflineEntheoGod
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Re: Help me find my error in WBS prep? [Re: tripdawg420]
    #26795394 - 06/30/20 07:38 PM (3 years, 6 months ago)

Ima make a different thread in a few days purely about my agar but for now I’d say this thread is pretty done. :smile: thanks for everyone’s input and help I look forward to the help in the future 😂.


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