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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome]
    #27418529 -

Looking to step up my sterile technique with my flow hood. Have mixed results with plates. Pouring is usually 100% success rate it’s more inoculation and transfers.

It’s mostly bacterial contam I get. I’m going to try an alcohol lamp so I’m not constantly going out of the flow to use the jet light. Also going to start wiping my scalpel handle with ISO more frequently as it’s not something I tend to do.

I think one of the things I may be doing wrong is this.. I find it easier to pick my Petri dish up to make transfers, so I have a little shot glass on my rack which I ISO first, and it’s mainly used to put my scalpel in after flaming it so I can sanitise my hands after touching the lighter. When I take a lid from a Petri dish, it gets placed carefully upside down onto the shot glass. But I’m guessing while working next to the lid in the dish, I’m potentially knocking contams into the lid?

Both the light and the bottle of ISO are out of flow when I work so I’m guessing this has a part to play. I don’t have a big flowhood.

Using my own PC’d swabs I have better success from print to agar. Other people’s swabs not so much. Bacterial disaster.

I need to understand my flowhood better and what I’m doing wrong, and what I can do change it.

I’m going to start doing some no pours along with my petris so I can start to have some results. I haven’t been spending much time in this hobby lately but I’m itching to dive back in.

Anything anybody can add that I might be missing that isn’t so obvious? Maybe the way the parafilm gets handled or if it is exposed to unclean air can contamination stick to it and then be inside your plate ? This is something I’ve wondered for a while.


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Being pissed on does not make you a real man.

...OR DOES IT?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: A.k.a] * 1
    #27418534 -

And thank you guys for treating me fairly lol,those groups roast you for your failures and never comment on good pics or questions or hail any success,I’m kinda glad I figured out how to use this website and I’m able to get in touch with the people who’s teks run through those groups like gold.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Guerrilla]
    #27418540 -

If it is only the next day and you can see healthy strands at all then it should live. You might have to clean it up but time will tell in that regard.

If you see life it should keep going. Good job.


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Question everything, it's very responsive.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Ukdude]
    #27418542 -

Ukdude said:
And thank you guys for treating me fairly lol,those groups roast you for your failures and never comment on good pics or questions or hail any success,I’m kinda glad I figured out how to use this website and I’m able to get in touch with the people who’s teks run through those groups like gold.


Well that sucks, fuck those guys lol.
Welcome to the shroomery.


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(You Know What Time It Is)
First they came for the communists, and I did not speak out because I was not a communist.
Then they came for the migrants, and I did not speak out because I was not a migrant.
Then they came for the queers, and I did not speak out because I was not queer.
Then they came for me, and there was nobody left to help me.
ABOLISH ICE. Free Palestine. Revolution or bust (A) . Profile pic credit goes to an awesome human
Be Gay, Do Crime 161 1312

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Nichrome]
    #27418544 -

Yeah I was expecting to have to do at least some cleanup. Just weirded me out how dark the samples look. Thanks!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Guerrilla]
    #27418618 -

Guerrilla said:
It’s mostly bacterial contam I get.


How confident are you in the sterility of your petris in the package? I only ask because I do many of the same things you're describing and have a pretty low contam rate.

I'm working in a carpeted, walk-in "closet" to boot. I rarely wear gloves and rather opt to wash my hands and use hand sanitizer before working. I do have a torch attachment on a camping propane tank in lieu of a torch lighter...

Edited by Lostkeys (08/07/21 07:20 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Guerrilla] * 1
    #27418626 -

This is how you make transfers in flow, the only difference is that you will be using a propane torch to sterilize your scalpel. Don't use an alcohol lamp, they suck balls. Sterilize your blade in flow, don't flip your lid, do what I do.



Do not hover your hands over the open dish, cool your blade on the bottom/downstream edge of your Petri.

I spin my dish so that the mycelium I'd like to select for transfer is on the bottom/downstream edge of the culture, this way you're not reaching over clean myc with your scalpel.

The source of your bacteria is not likely going to be from your agar, it's very easy to spot. Bacteria will more than likely be  a fault in grain preparation or incomplete sterilization.

Read through my clean spawn checklist in my signature.

Vent your pc.

Edited by Stipe-n Cap (08/07/21 07:22 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap]
    #27418638 -

Lostkeys said:
Guerrilla said:
It’s mostly bacterial contam I get.



How confident are you in the sterility of your petris in the package?


I don’t recall having issues with plates I pour and leave clean for a few weeks, so I don’t think that’s the problem, more that as much as I like to think I’m being clean and careful I’m probably doing something wrong.

p9hu7 said:
This is how you make transfers in flow, the only difference is that you will be using a propane torch to sterilize your scalpel. Don't use an alcohol lamp, they suck balls. Sterilize your blade in flow, don't flip your lid, do what I do.



Do not hover your hands over the open dish, cool your blade on the bottom/downstream edge of your Petri.

I spin my dish so that the mycelium I'd like to select for transfer is on the bottom/downstream edge of the culture, this way you're not reaching over clean myc with your scalpel.


Thanks dude that’s really helpful to me. I think I’ve just been taking too big of chunks, so I’ll do smaller as you do. My main stress was my plate was sliding over the place but as seen in your video it’s a normal inconvenience you just have to work around.

Also the comment about cooling and transferring at the bottom of the plate - I think that will be a game changer as I’ve definitely been leaning over the plate way too often. Big thank you.

What’s that tape used in that video?


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Being pissed on does not make you a real man.

...OR DOES IT?

Edited by Guerrilla (08/07/21 07:26 PM)

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap]
    #27418639 -

That's some deft handwork you got!

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap]
    #27418643 -

p9hu7 said:
This is how you make transfers in flow, the only difference is that you will be using a propane torch to sterilize your scalpel. Don't use an alcohol lamp, they suck balls. Sterilize your blade in flow, don't flip your lid, do what I do.



Do not hover your hands over the open dish, cool your blade on the bottom/downstream edge of your Petri.

I spin my dish so that the mycelium I'd like to select for transfer is on the bottom/downstream edge of the culture, this way you're not reaching over clean myc with your scalpel.

The source of your bacteria is not likely going to be from your agar, it's very easy to spot. Bacteria will more than likely be  a fault in grain preparation or incomplete sterilization.

Read through my clean spawn checklist in my signature.

Vent your pc.


Where did you find that little rack you're using? Closest thing I could find is about six inches tall. What is that sterilizer you're using?


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So heavy I fell through the earth...


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Re: AGAR ENVY! (Anything and All things agar!) [Re: Lostkeys]
    #27418654 -

I use cling wrap to wrap my plates.

As for the rack: I chop the legs off with a dremel tool, then put the foot caps back on:cool:

Definitely take small transfers, and I hate it when my plates slide! I'm going to buy some spray on rubber, or something. Hopefully that works.

The Sterilizer is a bactizapper, or a infrared Sterilizer for loops, scalpel, etc.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap]
    #27418667 -

Yeah, I looked that thing up... wish I hadn't.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap]
    #27418669 -

Thanks kindly brother. Do you have any tips/a description or a video on cutting and putting a plate to a grain jar?


--------------------
Being pissed on does not make you a real man.

...OR DOES IT?

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap]
    #27418670 -

p9hu7 said:
The Sterilizer is a bactizapper, or a infrared Sterilizer for loops, scalpel, etc.


I have a rubbing alcohol/propane torch induced scar that says I could use one of these.  Any suggestions on how to find one for a decent price?


--------------------
Why should we strive with cynic frown
To knock their fairy castles down?  ~ Eliza Cook

It's rather embarrassing to have given one's entire life to pondering the human predicament and to find that in the end one has little more to say than, 'Try to be a little kinder.' ~Aldous Huxley

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Guerrilla] * 1
    #27418673 -

It's the exact same process, except you're  using a larger wedge *1/4 dish, and then drop it in the loosened jar lid like I just did with the Petri.

I purchased from ebay.

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap]
    #27418696 -

Has anyone used the Chinese version?  There's a few of the unbranded 35mms going for around $150 right now.


--------------------
Why should we strive with cynic frown
To knock their fairy castles down?  ~ Eliza Cook

It's rather embarrassing to have given one's entire life to pondering the human predicament and to find that in the end one has little more to say than, 'Try to be a little kinder.' ~Aldous Huxley

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Stipe-n Cap]
    #27418699 -

I'd really love some opinions on some plates I made on the 5th...


I dropped a chunk of colonized BRF into this plate.


Also, the other day I posted a hella ugly plate with some drippy looking bacteria. I transfered (or tried) two spots on it.




I had another pretty ugly plate, too...





:facepalm: I know. I know. I'm new. I'd really appreciate any feedback. Thank you.


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My Grow Journal

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Paracosm]
    #27418722 -

those plates need to grow out some more before anyone can give a real opinion


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my FH build

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Re: AGAR ENVY! (Anything and All things agar!) [Re: monkey_accident]
    #27418725 -

Alright, thanks. I'll wait a couple more days and revisit it.


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My Grow Journal

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Re: AGAR ENVY! (Anything and All things agar!) [Re: Ukdude]
    #27418744 -

Ukdude said:
And thank you guys for treating me fairly lol,those groups roast you for your failures and never comment on good pics or questions or hail any success,I’m kinda glad I figured out how to use this website and I’m able to get in touch with the people who’s teks run through those groups like gold.


Shroomery on FB is fucking cancer. Def not my cup of tea over there :shrug:


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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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