StophJS said:

I have twice now tried making my own grainspawn with Bods easy AF oat tek. The oats are from a feed store and should be fine. The jars have plastic lids with two holes drilled for a SHIP and disc filter. I believe everything is sealed properly. Both times, all six jars ended up bacterial.

I used LC from one of the recommended sources. I did not test on agar, and I know the people here who do agar will stop me right there and want to say that my LC was contaminated. I think it's more likely there is something in the grain than both times in the LC. I do plan on testing on agar next go around but I believe I'll run into the same problem even if I confirm clean culture.

I'm just not sure how to proceed here. I don't want to overpay for prepared grain spawn. Should I try a different grain? I'm really not understanding how it is ending up that every single jar of these oats is bacterial.

Thanks.

Mr Piggy said:

I’m going to stop you right there.

You used untested vendor LC in grain, of course it went bacterial.  There is no “trusted source” besides your own work, period.

---

"Of course it went bacterial". My friend, that doesn't make sense... With all due respect, I do believe we should be able to float a hypothetical in these forums. It cannot assumed that the bacteria is not from the grainspawn or problems with the jars, and it's perfectly reasonable to discuss those possibilities, and frankly, probabilities.

---

Stuntbum said:

StophJS said:

I have twice now tried making my own grainspawn with Bods easy AF oat tek. The oats are from a feed store and should be fine. The jars have plastic lids with two holes drilled for a SHIP and disc filter. I believe everything is sealed properly. Both times, all six jars ended up bacterial.

I used LC from one of the recommended sources. I did not test on agar, and I know the people here who do agar will stop me right there and want to say that my LC was contaminated. I think it's more likely there is something in the grain than both times in the LC. I do plan on testing on agar next go around but I believe I'll run into the same problem even if I confirm clean culture.

I'm just not sure how to proceed here. I don't want to overpay for prepared grain spawn. Should I try a different grain? I'm really not understanding how it is ending up that every single jar of these oats is bacterial.

Thanks."Of course it went bacterial". My friend, that doesn't make sense... With all due respect, I do believe we should be able to float a hypothetical in these forums. It cannot assumed that the bacteria is not from the grainspawn or problems with the jars, and it's perfectly reasonable to discuss those possibilities, and frankly, probabilities.

---

How are you pressure cooking the grain? Whats your process for inoculation? If youre sure youre cleaning the grain properly, and youre sure that your LC is clean, where do you hypothesize the contaminates are entering the equation?

Is there something in the process youre not 100% sure of that you could narrow down and eliminate as a possible source of contamination?

---

GenesisCorrupted said:

You’re bringing mystery broth through the mail system and then wondering why it is dirty.

It is pretty notoriously well-known that liquid culture needs to be tested before it’s used, regardless of where it came from.

That’s why everyone is going to keep hammering you about agar.

It’s fun and easy.
Then make your own lc.

---

Smellyhobbit said:

Sterilize outside the box. Never put your flame inside the box.

I don’t use a false bottom or damp towel. I just go right on the countertop without issue.

A SAB is literal magic. The fact that you can use a flipped over tub with arm holes to do STERILE work is magic. We overthink it a little, imo

---

Arctic_Fox said:

GenesisCorrupted said:

You’re bringing mystery broth through the mail system and then wondering why it is dirty.

It is pretty notoriously well-known that liquid culture needs to be tested before it’s used, regardless of where it came from.

That’s why everyone is going to keep hammering you about agar.

It’s fun and easy.
Then make your own lc.

---

Why is LC so unreliable that you always need to test it on agar before using it? Is there no methodology to make LC with contamination rates below 10%?

---

ThurmanMurman said:

Im gonna go change my diaper now that I'm done being a baby.

---

Arctic_Fox said:

GenesisCorrupted said:

You’re bringing mystery broth through the mail system and then wondering why it is dirty.

It is pretty notoriously well-known that liquid culture needs to be tested before it’s used, regardless of where it came from.

That’s why everyone is going to keep hammering you about agar.

It’s fun and easy.
Then make your own lc.

---

Why is LC so unreliable that you always need to test it on agar before using it? Is there no methodology to make LC with contamination rates below 10%?

---

ThurmanMurman said:

Smellyhobbit said:

Sterilize outside the box. Never put your flame inside the box.

I don’t use a false bottom or damp towel. I just go right on the countertop without issue.

A SAB is literal magic. The fact that you can use a flipped over tub with arm holes to do STERILE work is magic. We overthink it a little, imo

---

I would actually really like to be able to get away from the false bottom. Im incredibly fat fingered, between trying not to bump the box just within the arm holes and trying to make sure I grab the bottom half of a plastic petri while its in a stack so as not to expose the inside by accidentally grabbing the lid, and then "Oh! You're getting pretty close to ceiling too bud, so keep an eye on that!" I feel like I'm playing the game Operation, except when I screw up, I've just pissed away days or weeks. I dont even feel like its something I can improve through practice either - I've never been one for delicate or precision movements. My favorite type of work is more in the wheelhouse of a jackhammer, know what I mean? I think I'll limp through it fine, but an FFU is definitely at the top of my list. Sterile movements are one thing, but trying to do it all while not bumping into anything is another. Thanks for listening to me vent about how the box stresses me out. Im gonna go change my diaper now that I'm done being a baby.

---

My hands were shaking like a leaf while I was injecting my batch that turned out well. It really sounds like your SAB is too cluttered or maybe your arm holes are too small or both. You can make them pretty big and it can actually work out better then small holes. But yeah with your jars straight on the counter that may help your space issues significantly. Idk if this will help but when I felt like I waited too long trying to get into the injection holes I just flame sterilized and tried again.

Also I got some really good advice to do a practice run first. I practiced maybe 3 times. Just keep the needle cap on (I didn't even put the needle on but the expert said you can as long as the caps on) wash your hands and practice a bit.

Hope this helps. I know you can do this

StophJS said:

Stuntbum said:

StophJS said:

I have twice now tried making my own grainspawn with Bods easy AF oat tek. The oats are from a feed store and should be fine. The jars have plastic lids with two holes drilled for a SHIP and disc filter. I believe everything is sealed properly. Both times, all six jars ended up bacterial.

I used LC from one of the recommended sources. I did not test on agar, and I know the people here who do agar will stop me right there and want to say that my LC was contaminated. I think it's more likely there is something in the grain than both times in the LC. I do plan on testing on agar next go around but I believe I'll run into the same problem even if I confirm clean culture.

I'm just not sure how to proceed here. I don't want to overpay for prepared grain spawn. Should I try a different grain? I'm really not understanding how it is ending up that every single jar of these oats is bacterial.

Thanks."Of course it went bacterial". My friend, that doesn't make sense... With all due respect, I do believe we should be able to float a hypothetical in these forums. It cannot assumed that the bacteria is not from the grainspawn or problems with the jars, and it's perfectly reasonable to discuss those possibilities, and frankly, probabilities.

---

How are you pressure cooking the grain? Whats your process for inoculation? If youre sure youre cleaning the grain properly, and youre sure that your LC is clean, where do you hypothesize the contaminates are entering the equation?

Is there something in the process youre not 100% sure of that you could narrow down and eliminate as a possible source of contamination?

---

I think a reasonable next step is just to test on agar and keep all other variables exactly the same. I'm PCing for two hours at 17, venting properly and all that. I thought maybe my lids, or the oats are just somehow still harboring the bacteria. At least if I test and use wedges next time as opposed to injecting and it still goes south I'll know for sure which end it's on.

---

Coconutz said:

Hey guys, can you stack jars on top of each other when doing a steam bath for PFtek?

---

StophJS said:

GenesisCorrupted said:

Stopping you right there.
Why make a post about you not wanting to use the very easy (less than a dollar) method.

Buying grain or lc sent through the mail wouldn’t make me feel better.
I want to see a pic of these “fine” lids.

---

Thought I did a good job with them, but perhaps not. As I said I will use agar next time. But I don't want the same thing to happen, and I suspect it will.

---

Quote:

ThurmanMurman said:

Smellyhobbit said:

Sterilize outside the box. Never put your flame inside the box.

I don’t use a false bottom or damp towel. I just go right on the countertop without issue.

A SAB is literal magic. The fact that you can use a flipped over tub with arm holes to do STERILE work is magic. We overthink it a little, imo

---

I would actually really like to be able to get away from the false bottom. Im incredibly fat fingered, between trying not to bump the box just within the arm holes and trying to make sure I grab the bottom half of a plastic petri while its in a stack so as not to expose the inside by accidentally grabbing the lid, and then "Oh! You're getting pretty close to ceiling too bud, so keep an eye on that!" I feel like I'm playing the game Operation, except when I screw up, I've just pissed away days or weeks. I dont even feel like its something I can improve through practice either - I've never been one for delicate or precision movements. My favorite type of work is more in the wheelhouse of a jackhammer, know what I mean? I think I'll limp through it fine, but an FFU is definitely at the top of my list. Sterile movements are one thing, but trying to do it all while not bumping into anything is another. Thanks for listening to me vent about how the box stresses me out. Im gonna go change my diaper now that I'm done being a baby.

---

I have the same issue to a degree, I’m an extremely shakey person and always have been. I will say that over time it’s gotten a lot more casual and comfortable and with that I’ve gotten much less shakey.

I don’t fear contam anymore, and lots of times I may not bother closing the room door or waiting a full 30 after putting stuff in the SAB. I’ll put some plates in, wait 2 minutes or so, and do transfers without spraying anything and they’ll be clean. Every now and again a plate gets some bacteria but I feel like the point of agar is to easily be able to transfer away from contam. So it’s much less scary now and I’m confident that if I see any contam I can transfer away from it easily.

I found that using a wire rack was the #1 factor of reducing contam besides just regular sterile technique for me personally.

To me it’s a bit like gaming. I used to play games and hide and be scared and then boom I’m dead, now I just run in with confidence and boom I’m usually the one killing the one hiding lol.

SAB work is like that to me

Seriously_trippin said:

ThurmanMurman said:

Smellyhobbit said:

Sterilize outside the box. Never put your flame inside the box.

I don’t use a false bottom or damp towel. I just go right on the countertop without issue.

A SAB is literal magic. The fact that you can use a flipped over tub with arm holes to do STERILE work is magic. We overthink it a little, imo

---

I would actually really like to be able to get away from the false bottom. Im incredibly fat fingered, between trying not to bump the box just within the arm holes and trying to make sure I grab the bottom half of a plastic petri while its in a stack so as not to expose the inside by accidentally grabbing the lid, and then "Oh! You're getting pretty close to ceiling too bud, so keep an eye on that!" I feel like I'm playing the game Operation, except when I screw up, I've just pissed away days or weeks. I dont even feel like its something I can improve through practice either - I've never been one for delicate or precision movements. My favorite type of work is more in the wheelhouse of a jackhammer, know what I mean? I think I'll limp through it fine, but an FFU is definitely at the top of my list. Sterile movements are one thing, but trying to do it all while not bumping into anything is another. Thanks for listening to me vent about how the box stresses me out. Im gonna go change my diaper now that I'm done being a baby.

---

My hands were shaking like a leaf while I was injecting my batch that turned out well. It really sounds like your SAB is too cluttered or maybe your arm holes are too small or both. You can make them pretty big and it can actually work out better then small holes. But yeah with your jars straight on the counter that may help your space issues significantly. Idk if this will help but when I felt like I waited too long trying to get into the injection holes I just flame sterilized and tried again.

Also I got some really good advice to do a practice run first. I practiced maybe 3 times. Just keep the needle cap on (I didn't even put the needle on but the expert said you can as long as the caps on) wash your hands and practice a bit.

Hope this helps. I know you can do this

---

Dkeppel said:

Same here, adding a wire rack and having confidence in my technique has been crucial.

I used to let plates sit around for a couple of week before use, and drop a single wedge into grain with uncertainty.

Now I'll use plates straight away, and I'll cut a plate into a tic-tac-toe pattern, then use my scalpel to dislodge them all into the grain jar like Beef does in the below link, except I keep an empty jar in the SAB when using 2-piece lids to rest the lid on while dislodging.

https://www.shroomery.org/forums/showflat.php/Number/28744657

When I started doing the tic-tac-toe technique a couple months ago, I thought there was no fucken way this technique would be success without a LFH/FFU, but of 30 odd jars inoculated this way, not a single jar has contaminated yet.

---