37 replies to this topic

[jrh]

Is it normal to get any contams? I think out of 40 ketchup cups I made, 2 were contaminated when I went to use them, and out of 10 petri dishes, 1 was contaminated. It was my 2nd time pouring ketchup cups and my 1st time pouring petri dishes.

[Auhron]

It happens. my agar experience involves a lot of contamination.   , It takes some practice to get your sterile technique drawn in.  though, if you keep doing it at those numbers you have nothing to worry about.

[rockyfungus]

It's a process. Did you try to inoculate the ketchup cups? Or were they just sitting out.

Im comfortable in my process, but when I change any part. I'll leave a few plates untouched to see if I messed up pouring. Or with the agar. If In a few days to a week nothing shows up, your technique making agar is fine.

Now you need to check what is happening as you reopen the cups, or go to inoculate them.

[jrh]

Yes, I ended up using most of the ketchup cups (still have 8 left). They were just sitting around, and when I went to use them a couple had pin mold. Of the petri dishes, one had a bacterial growth when I did some transfers last night.

 

BTW, is there an "approved" way of getting spore syringe to agar? Even if I do a single drop, it ends up going everywhere. Maybe put a little into a ketchup cup and use a swab or loop to put some down just in the center?

[rockyfungus]

I tend to do a small drop and swirl my petri dish. I've also used an innoculation loop and sterilized qtips.

Loop/swab is recommended.

My problem with ketchup cups, the lids will warp and suck some unsterile air in.

[jrh]

My problem with ketchup cups, the lids will warp and suck some unsterile air in.

 

I've been pouring at around 120 degrees f and have been lucky so far (although I had a lot of condensation). I think I'm going to use cups primarily to get something growing and isolate it, and when I have something I think might be good, transfer it to petri dish.

[ItBeBasidia]

@jrh
The type of soufflet cup matters too. Make sure the brand you get has completely sealed cellophane packaging. Some are stapled shut and those are more apt to contam. Ive had great success with the Walmart brand ones.

@PJammer24
The bunsen and flame updraft definitely work. It's the method used in many universities when the flow hoods are taken up. The oven updraft on the other hand has to have the heat set pretty high as while as deliberate movement and environmental awareness to be effective. The key is in creating an updraft. Work must be done above/near the cracked oven door, not horizontally with the door wide open in a horizontal draft. I personally prefer a butane torch or a single propane camp stove. For the rambling cultivator that has to worry about prying eyes(uni dorm residents, Mil personnel, business travelers, etc.) These sterile technique hacks will keep em growing.

Here's a pic of how the oven door should be.
If you put an incense stick near, it'll push the smoke up and out away from the "cone of protection". Don't mind my dirty stove top I just cooked dinner.

Edited by ItBeBasidia, 17 December 2020 - 09:25 PM.

[Alpoehi]

... For the rambling cultivator that has to worry about prying eyes(uni dorm residents, Mil personnel, business travelers, etc.) These sterile technique hacks will keep em growing.

 

Oh man that's me 

[jrh]

@jrh
The type of soufflet cup matters too. Make sure the brand you get has completely sealed cellophane packaging. Some are stapled shut and those are more apt to contam. Ive had great success with the Walmart brand ones.

 

Thanks! Someone else tipped me off about the walmart brand cups. Now I've just got to learn to get rid of the condensation. Maybe a grow mat on top as they're cooling, or after I've packaged them 4 to a ziploc sandwich bag.

[jrh]

Nutritional yeast question for you all. I bought a little bag of nutritional yeast because Frank's list of agar recipes suggested that LME + nutritional yeast was super duper stuff. My nutritional yeast looks like fish food flakes. I can see from looking in my agar dishes that mycelium comes across a flake of nutritional yeast and really thrives on that spot.

 

The question part is do you think it would be advantageous to blitz my nutritional yeast flakes into a powder? Or would it all just clump and become unusable?

[Auhron]

It will be fine. powder it up. It will distribute more even through your agar that way.

[jrh]

I just powdered it -- now I feel like the agar dishes I made yesterday are inferior. 

 

Anyways, I also made up 2 LC jars, and I added a scant pinch of the nutritional yeast to those. I figure it will sink, but so will cube spores and mycelium, so maybe it'll be a nom nom match made in heaven. Or maybe it'll just cloud up the LC and I'll have to throw them away. I should figure that out in a week or two.

Edited by jrh, 18 December 2020 - 07:29 AM.

[PJammer24]

@jrh
The type of soufflet cup matters too. Make sure the brand you get has completely sealed cellophane packaging. Some are stapled shut and those are more apt to contam. Ive had great success with the Walmart brand ones.

@PJammer24
The bunsen and flame updraft definitely work. It's the method used in many universities when the flow hoods are taken up. The oven updraft on the other hand has to have the heat set pretty high as while as deliberate movement and environmental awareness to be effective. The key is in creating an updraft. Work must be done above/near the cracked oven door, not horizontally with the door wide open in a horizontal draft. I personally prefer a butane torch or a single propane camp stove. For the rambling cultivator that has to worry about prying eyes(uni dorm residents, Mil personnel, business travelers, etc.) These sterile technique hacks will keep em growing.

 

 

 

I understand the concept, I am just pretty sure that there have been some tests done on here and that the oven was proven ineffective... I don't see much need for it anyway... It is pretty easy to fashion an SAB for cheap and the success rates for SABs is pretty darn good.

[ItBeBasidia]

That's why it's in an alternative thread. Some people need ways to grow that are oddball. For instance, people that get room inspections might prefer not to have to explain to the inspector, commander, or dorm chief what the tote/cardboard box with armholes, alcohol, and exacto knife inside it is.

It's also good to have all methods of cultivation documented so we don't become a single way to grow type of site.

I'm hoping every different way about cultivation finds its way to this thread for people that may need an improv solution.

It'd be cool if people also posted their trials and success rates with some of these methods or links to it on this thread.

Edited by ItBeBasidia, 18 December 2020 - 01:17 PM.

[ItBeBasidia]

Here's some awesome work being done to make flow hoods:

https://mycotopia.ne...08724-Scrap-LFH

https://mycotopia.ne...ask-me-anything

[ItBeBasidia]

Experimenting with low cost flowhood solutions.

So far $10 for tote, $25 for filter, and $3 for air pump (thrift shop).

Already anticipating changing the air pump for a hydroponics ventilation fan because I doubt it'll push enough air. It's loud too.

[bezevo]

no  cllue is this thrufty laminar works but GORDO TEK guy usually has good vids

 

https://www.patreon....ource=post_link

[ItBeBasidia]

Put a new fan that's really quiet and tested on agar on April 1st. So far everything is clean. Some GT spores i did along the with the blank agar germinated too with no signs of contam. 2nd pic is how I store it when not in use, except for I also put a sandwich bag over the fan.