Home | Community | Message Board

MRCA Tyroler Gluckspilze
This site includes paid links. Please support our sponsors.


Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Shop: PhytoExtractum Buy Bali Kratom Powder   North Spore Injection Grain Bag   Kraken Kratom Red Vein Kratom   Original Sensible Seeds Autoflowering Cannabis Seeds   Left Coast Kratom Buy Kratom Capsules   Unfolding Nature Unfolding Nature: Being in the Implicate Order   Mushroom-Hut Liquid Cultures

Jump to first unread post Pages: 1 | 2 | 3 | Next >  [ show all ]
OfflineEntheoGod
Entheo
Male User Gallery


Registered: 03/06/15
Posts: 648
Last seen: 8 months, 15 days
Help me find my error in WBS prep?
    #26785017 - 06/26/20 03:00 PM (3 years, 7 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)


Extras: Filter Print Post Top
OfflineEntheoGod
Entheo
Male User Gallery


Registered: 03/06/15
Posts: 648
Last seen: 8 months, 15 days
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785127 - 06/26/20 04:00 PM (3 years, 7 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.


Extras: Filter Print Post Top
Offlineredhandmat
Dude


Registered: 05/09/19
Posts: 463
Last seen: 2 years, 8 months
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785182 - 06/26/20 04:33 PM (3 years, 7 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)


Extras: Filter Print Post Top
OfflineEntheoGod
Entheo
Male User Gallery


Registered: 03/06/15
Posts: 648
Last seen: 8 months, 15 days
Re: Help me find my error in WBS prep? [Re: redhandmat]
    #26785205 - 06/26/20 04:44 PM (3 years, 7 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)


Extras: Filter Print Post Top
Offlineredhandmat
Dude


Registered: 05/09/19
Posts: 463
Last seen: 2 years, 8 months
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785252 - 06/26/20 05:05 PM (3 years, 7 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.


Extras: Filter Print Post Top
OfflineEntheoGod
Entheo
Male User Gallery


Registered: 03/06/15
Posts: 648
Last seen: 8 months, 15 days
Re: Help me find my error in WBS prep? [Re: redhandmat]
    #26785282 - 06/26/20 05:15 PM (3 years, 7 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! 😊


Extras: Filter Print Post Top
Offlineredhandmat
Dude


Registered: 05/09/19
Posts: 463
Last seen: 2 years, 8 months
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785298 - 06/26/20 05:22 PM (3 years, 7 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.


Extras: Filter Print Post Top
OfflineEntheoGod
Entheo
Male User Gallery


Registered: 03/06/15
Posts: 648
Last seen: 8 months, 15 days
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785301 - 06/26/20 05:25 PM (3 years, 7 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)!


Extras: Filter Print Post Top
OfflineEntheoGod
Entheo
Male User Gallery


Registered: 03/06/15
Posts: 648
Last seen: 8 months, 15 days
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785361 - 06/26/20 05:56 PM (3 years, 7 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.


Extras: Filter Print Post Top
Offlinelateforthafuture
fanatic
 User Gallery

Registered: 03/26/07
Posts: 471
Last seen: 3 years, 4 months
Re: Help me find my error in WBS prep? [Re: redhandmat]
    #26785436 - 06/26/20 06:32 PM (3 years, 7 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)


Extras: Filter Print Post Top
InvisibleFriedEggS
I'm a teapot


Registered: 09/22/14
Posts: 2,537
Loc: Taiwan
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785437 - 06/26/20 06:33 PM (3 years, 7 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.


Extras: Filter Print Post Top
OfflineEntheoGod
Entheo
Male User Gallery


Registered: 03/06/15
Posts: 648
Last seen: 8 months, 15 days
Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26785458 - 06/26/20 06:41 PM (3 years, 7 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)


Extras: Filter Print Post Top
Offlinelateforthafuture
fanatic
 User Gallery

Registered: 03/26/07
Posts: 471
Last seen: 3 years, 4 months
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785463 - 06/26/20 06:47 PM (3 years, 7 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
:superbanana::mjdance::dancer:


Edited by lateforthafuture (06/26/20 06:49 PM)


Extras: Filter Print Post Top
InvisibleFriedEggS
I'm a teapot


Registered: 09/22/14
Posts: 2,537
Loc: Taiwan
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785468 - 06/26/20 06:50 PM (3 years, 7 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:


Extras: Filter Print Post Top
OfflineEntheoGod
Entheo
Male User Gallery


Registered: 03/06/15
Posts: 648
Last seen: 8 months, 15 days
Re: Help me find my error in WBS prep? [Re: lateforthafuture]
    #26785486 - 06/26/20 06:57 PM (3 years, 7 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)


Extras: Filter Print Post Top
Offlinelateforthafuture
fanatic
 User Gallery

Registered: 03/26/07
Posts: 471
Last seen: 3 years, 4 months
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785508 - 06/26/20 07:05 PM (3 years, 7 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.


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


Extras: Filter Print Post Top
OfflineEntheoGod
Entheo
Male User Gallery


Registered: 03/06/15
Posts: 648
Last seen: 8 months, 15 days
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785511 - 06/26/20 07:07 PM (3 years, 7 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)


Extras: Filter Print Post Top
InvisibleFriedEggS
I'm a teapot


Registered: 09/22/14
Posts: 2,537
Loc: Taiwan
Re: Help me find my error in WBS prep? [Re: EntheoGod]
    #26785532 - 06/26/20 07:15 PM (3 years, 7 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.


--------------------

(Yes, the egg is real)

How to post pics


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


Extras: Filter Print Post Top
InvisibleFriedEggS
I'm a teapot


Registered: 09/22/14
Posts: 2,537
Loc: Taiwan
Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26785537 - 06/26/20 07:17 PM (3 years, 7 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.


--------------------

(Yes, the egg is real)

How to post pics


Extras: Filter Print Post Top
Offlinelateforthafuture
fanatic
 User Gallery

Registered: 03/26/07
Posts: 471
Last seen: 3 years, 4 months
Re: Help me find my error in WBS prep? [Re: FriedEgg]
    #26785547 - 06/26/20 07:20 PM (3 years, 7 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.


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


Extras: Filter Print Post Top
Jump to top Pages: 1 | 2 | 3 | Next >  [ show all ]

Shop: PhytoExtractum Buy Bali Kratom Powder   North Spore Injection Grain Bag   Kraken Kratom Red Vein Kratom   Original Sensible Seeds Autoflowering Cannabis Seeds   Left Coast Kratom Buy Kratom Capsules   Unfolding Nature Unfolding Nature: Being in the Implicate Order   Mushroom-Hut Liquid Cultures


Similar ThreadsPosterViewsRepliesLast post
* Foo Man's WBS Method
( 1 2 3 4 ... 9 10 all )
FooMan 119,239 180 02/22/23 01:03 PM
by johnukguy
* WBS Prep hydr0420 1,776 3 06/15/04 08:25 PM
by Gr0wer
* WBS Prep. Question manto2 2,742 15 12/08/04 09:49 AM
by MushroomFriend
* Soak WBS mrhowdy 4,247 4 01/22/05 08:14 PM
by mrhowdy
* wbs wetness and brand? reflectedlight 1,496 7 05/12/05 08:45 AM
by agar
* WBS coming out of PC dry and gooey jamman 2,320 12 01/23/06 06:54 AM
by doc34
* WBS mixed 2:1 w/ water? jtk 734 6 07/01/05 03:08 PM
by korins
* - ShroomGuy86 584 8 03/08/05 07:59 PM
by LengthyFellow

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: Shroomism, george castanza, RogerRabbit, veggie, mushboy, fahtster, LogicaL Chaos, 13shrooms, Stipe-n Cap, Pastywhyte, bodhisatta, Tormato, Land Trout, A.k.a
645 topic views. 36 members, 182 guests and 41 web crawlers are browsing this forum.
[ Show Images Only | Sort by Score | Print Topic ]
Search this thread:

Copyright 1997-2024 Mind Media. Some rights reserved.

Generated in 0.025 seconds spending 0.008 seconds on 14 queries.