KauaiOrca said:

Anyone ever done this?

I was near the end of a spore syringe (purchased from vendor) and I had some clean LME LC (hadn't been inoculated). So I decided to leave about 1ml of spore solution in the syringe and I flamed the needle and drew up 5ml of the LME liquid into the syringe and the rest with air from inside the LME jar then capped the syringe so that the syringe has 5ml of LC liquid and 5ml of air.

A week later, it was colonizing really well in an airtight syringe container! No visual signs of contams.

I thought it would stall out because there was no FAE at all, but it didn't... grew out really healthy and tested in on a few small pucks that grew out clean. Didn't test on agar and will, but nonetheless an interesting way to make a small syringe of LC.

I might try this at some point with cloning... Have 15ml LME in a 30ml syringe, poke a fruit and suck in hopefully some live mycelium and see what happens. Would be a pretty simple way to make clean LC with very low contam risk. Could even PC the syringe half filled with LME liquid and a needle so it's one easy step.

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Bitchfist said:

The plan was to just fruit in the bag. Cut the top open and leave it half sealed and give it FAE 3-4 times a day. It still has some small areas on the top that are not completely colonized. Should I let it wait about her couple days?

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KauaiOrca said:

Anyone ever done this?

I was near the end of a spore syringe (purchased from vendor) and I had some clean LME LC (hadn't been inoculated). So I decided to leave about 1ml of spore solution in the syringe and I flamed the needle and drew up 5ml of the LME liquid into the syringe and the rest with air from inside the LME jar then capped the syringe so that the syringe has 5ml of LC liquid and 5ml of air.

A week later, it was colonizing really well in an airtight syringe container! No visual signs of contams.

I thought it would stall out because there was no FAE at all, but it didn't... grew out really healthy and tested in on a few small pucks that grew out clean. Didn't test on agar and will, but nonetheless an interesting way to make a small syringe of LC.

I might try this at some point with cloning... Have 15ml LME in a 30ml syringe, poke a fruit and suck in hopefully some live mycelium and see what happens. Would be a pretty simple way to make clean LC with very low contam risk. Could even PC the syringe half filled with LME liquid and a needle so it's one easy step.

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LadysKnight said:

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KauaiOrca said:

LadysKnight said:

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No matter how few, spores are not sterile

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Bitchfist said:

This was one of the grow bags that has the grain on the bottom where the liquid culture is injected. That as already been mixed into the soil part of the bag (top layer). I’m a first time grower so I haven’t tried anything else.

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LadysKnight said:

KauaiOrca said:

LadysKnight said:

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No matter how few, spores are not sterile

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I'll grow out a few pucks next week and we'll see.

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LadysKnight said:

With the selective puck step, I can see that working, but spore syringe + LC broth is still the worst way to start. There is bacteria in that syringe of warm sugar water multiplying much faster than the spores can germinate. You should actually get better results if sterile water was added to the syringe instead. Maybe rgs is more bacteria resistant than grain.

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KauaiOrca said:

LadysKnight said:

With the selective puck step, I can see that working, but spore syringe + LC broth is still the worst way to start. There is bacteria in that syringe of warm sugar water multiplying much faster than the spores can germinate. You should actually get better results if sterile water was added to the syringe instead. Maybe rgs is more bacteria resistant than grain.

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I agree with that and don't usually do this.  This was more of a "what the hell" kind of test. Totally spur of the moment as I had a small about of spore solution and a jar of sterilize LME liquid so I just stuck it in there to see what would happen.

I typically use clone tissue to start new pucks.

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Ain't a damn thing wrong with trying stuff. Can be educational. I think you're showing RGS is bacteria resistant.

LadysKnight said:

KauaiOrca said:

LadysKnight said:

With the selective puck step, I can see that working, but spore syringe + LC broth is still the worst way to start. There is bacteria in that syringe of warm sugar water multiplying much faster than the spores can germinate. You should actually get better results if sterile water was added to the syringe instead. Maybe rgs is more bacteria resistant than grain.

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I agree with that and don't usually do this.  This was more of a "what the hell" kind of test. Totally spur of the moment as I had a small about of spore solution and a jar of sterilize LME liquid so I just stuck it in there to see what would happen.

I typically use clone tissue to start new pucks.

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Ain't a damn thing wrong with trying stuff. Can be educational. I think you're showing RGS is bacteria resistant.

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ThurmanMurman said:

LadysKnight said:

KauaiOrca said:

LadysKnight said:

With the selective puck step, I can see that working, but spore syringe + LC broth is still the worst way to start. There is bacteria in that syringe of warm sugar water multiplying much faster than the spores can germinate. You should actually get better results if sterile water was added to the syringe instead. Maybe rgs is more bacteria resistant than grain.

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I agree with that and don't usually do this.  This was more of a "what the hell" kind of test. Totally spur of the moment as I had a small about of spore solution and a jar of sterilize LME liquid so I just stuck it in there to see what would happen.

I typically use clone tissue to start new pucks.

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Ain't a damn thing wrong with trying stuff. Can be educational. I think you're showing RGS is bacteria resistant.

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Not to butt in, but to piggy back on "trying stuff", this is 100% how I learn best. Even if someone tells me it will fail, if it's not too much of a loss to see it for myself, I'm not against doing it anyway just for learning purposes. Knowing =/= understanding. 🤣

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How do other people do it?

LadysKnight said:

KauaiOrca said:

LadysKnight said:

With the selective puck step, I can see that working, but spore syringe + LC broth is still the worst way to start. There is bacteria in that syringe of warm sugar water multiplying much faster than the spores can germinate. You should actually get better results if sterile water was added to the syringe instead. Maybe rgs is more bacteria resistant than grain.

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I agree with that and don't usually do this.  This was more of a "what the hell" kind of test. Totally spur of the moment as I had a small about of spore solution and a jar of sterilize LME liquid so I just stuck it in there to see what would happen.

I typically use clone tissue to start new pucks.

---

Ain't a damn thing wrong with trying stuff. Can be educational. I think you're showing RGS is bacteria resistant.

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