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 SpaceBaby
alchemist
 
Registered: 11/01/20
Posts: 2,030
Loc: SPACE
Last seen: 7 months, 1 day
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 Re: Recognizing and dealing with contamination  [Re: B H O]
#27383597 - 07/11/21 08:02 PM (2 years, 9 months ago) |
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Quote:
B H O said:
So I made transfers from a really nice look rhizo plate and then shook the jars. During the transfer I felt like something was off about the agar plate.
When I pushed my hole punch into the plate, it felt more gel-like and soft than how my plates normally feel which is more hard/solid. When I tried to lift up on the hole punched agar with my scalpel, the agar stayed in place and only the mycelium was coming up with it. I ended up growing through with inoculating the rest of the jars anyway and the jars above were the result.
(Agar recipe used 10g Agar 9g LME 500ml water. I normally do 400ml water so maybe that’s why my texture felt off?)
I can’t tell whether it’s contaminated or the mycelium reverted into tomentose growth. If it is the later, I was always under the assumption that tomentose growth is a little slower how ever most of these jars are at 90% complete in like 4 days.
I guess I can try to fruit in a shoebox and see what happens, but wanted to see what you guys think.
   
I've done well with sketchy spawn in shoeboxes. Well being a relative term.
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SpaceBaby
SPACEBABY'S LAGM22 THREAD
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Me as a cube
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 Smartattack
C'mon man
Registered: 12/21/18
Posts: 3,779
Loc: A thought
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Re: Recognizing and dealing with contamination [Re: SpaceBaby]
#27383663 - 07/11/21 08:57 PM (2 years, 9 months ago) |
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Plate looks nice, jars look like garb though.
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* Smarts videos 
*  Planet of the APES
   
I'm a fungal white supremacist.
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B H O
Registered: 03/17/13
Posts: 675
Last seen: 1 month, 7 days
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Re: Recognizing and dealing with contamination [Re: Smartattack]
#27383677 - 07/11/21 09:09 PM (2 years, 9 months ago) |
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Quote:
Smartattack said:
Plate looks nice, jars look like garb though.
granted, I haven't fruited a jar in 6 years.. these looked like the ones that would give me trichoderma back in them days. I held off from throwing them away and they look a little better now. I hope that's the case. I'll keep ya updated I suppose. I have more prepped coir than I know what to do with rn anyway...
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Luminous7
Registered: 01/26/16
Posts: 1,428
Last seen: 13 hours, 18 minutes
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Re: Recognizing and dealing with contamination [Re: Smartattack]
#27383679 - 07/11/21 09:10 PM (2 years, 9 months ago) |
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Quote:
Smartattack said:
Plate looks nice, jars look like garb though.
Ya im suprised you had issues from that plate.
Could have been grain prep / sterilization went wrong somehow. Or sterile tek when knocking jars was off.
Plate looks mint.
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B H O
Registered: 03/17/13
Posts: 675
Last seen: 1 month, 7 days
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Re: Recognizing and dealing with contamination [Re: Luminous7]
#27383701 - 07/11/21 09:37 PM (2 years, 9 months ago) |
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Same!! It could be my grain prep but I used the same WBS jars to inoculate some Z strain which is doing alright. It could be from inoculation. Doing transfers onto grain sucks in a SAB 😂 I used to Lysol the shit out of a bathroom back in the day without problems now I feel like I’m getting more working in a SAB.
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gone-pear-shaped
Stranger than fiction
Registered: 10/30/17
Posts: 822
Last seen: 8 months, 2 days
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Re: Recognizing and dealing with contamination [Re: B H O]
#27383763 - 07/11/21 11:26 PM (2 years, 9 months ago) |
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Related to this topic, when a wedge sticks to the scalpel so you tap the scalpel on the jar rim to release it, is that likely to cause contamination? The neck of the scalpel was flamed (dipped in alcohol and lit) but not heated red. (The blade was heated dull red, though.)
If touching the jar is bad, how do you get the agar to release?
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LadysKnight
Hello Ladies
Registered: 10/09/15
Posts: 1,898
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I try to keep any vibration to a minimum. It's fine to use the rim of the jar to release the agar, only the agar wedge and the tip of the blade should pass the rim.
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B H O
Registered: 03/17/13
Posts: 675
Last seen: 1 month, 7 days
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Re: Recognizing and dealing with contamination [Re: LadysKnight]
#27383970 - 07/12/21 06:55 AM (2 years, 9 months ago) |
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That makes sense! Totally don’t know if I did or not. Definitely will be more aware of that now! Thanks 🙏
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gone-pear-shaped
Stranger than fiction
Registered: 10/30/17
Posts: 822
Last seen: 8 months, 2 days
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Re: Recognizing and dealing with contamination [Re: LadysKnight]
#27383977 - 07/12/21 07:03 AM (2 years, 9 months ago) |
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Quote:
LadysKnight said:
I try to keep any vibration to a minimum. It's fine to use the rim of the jar to release the agar, only the agar wedge and the tip of the blade should pass the rim.
Hmm... I put in most of the plate last time, so I piled it up on the scalpel. It touched beyond the blade. That probably wasn't smart, but I don't know how else to do it. Do you just add wedges one at a time?
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LadysKnight
Hello Ladies
Registered: 10/09/15
Posts: 1,898
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I use one wedge per jar. I inoculate 10 jars with one 100mm plate. I haven't seen enough of an improvement to use more than that. Also my PC does 10 quarts at a time. I only have to deal with one donor plate so less in and out, etc.
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bagga
Day Tripper
Registered: 05/19/21
Posts: 334
Last seen: 10 months, 18 days
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Agar stuck on the blade drives me nuts. Needs a tek 😂
Another member here made a scraper with a flame sterilized stainless paperclip (any thin wire that can take the heat would do).
It works for me, a lot less swearing.
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Luminous7
Registered: 01/26/16
Posts: 1,428
Last seen: 13 hours, 18 minutes
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Re: Recognizing and dealing with contamination [Re: B H O]
#27384328 - 07/12/21 11:22 AM (2 years, 8 months ago) |
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Quote:
B H O said:
Same!! It could be my grain prep but I used the same WBS jars to inoculate some Z strain which is doing alright. It could be from inoculation. Doing transfers onto grain sucks in a SAB 😂 I used to Lysol the shit out of a bathroom back in the day without problems now I feel like I’m getting more working in a SAB.
The principals of still air are more important than sterility, remember that.
Close all windows, turn off the vents in the house and anything that may cause draft.
Wear clean clothes , scrub forearms with soap, all that prep has helped me have good success in lab work.
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Luminous7
Registered: 01/26/16
Posts: 1,428
Last seen: 13 hours, 18 minutes
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Re: Recognizing and dealing with contamination [Re: Luminous7]
#27384330 - 07/12/21 11:24 AM (2 years, 8 months ago) |
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I always use tweezers and a scalple. Its 1 more thing to flame sterilize but i prefer the ease of work flow.
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NightPuma1
Stranger
Registered: 03/28/16
Posts: 310
Last seen: 2 years, 2 months
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Re: Recognizing and dealing with contamination [Re: Luminous7]
#27384491 - 07/12/21 01:28 PM (2 years, 8 months ago) |
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You should put the picture from my latest post in your guide as well 👌🏻
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laruz
Stranger
Registered: 06/01/21
Posts: 27
Last seen: 8 months, 6 days
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Re: Recognizing and dealing with contamination [Re: NightPuma1]
#27385586 - 07/13/21 08:57 AM (2 years, 8 months ago) |
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Hi guys ! now iam at my third attempt and i think i finally colonized my first jars. just to be sure please check this photo.. cause the mycelium seems really fluffy and iam not sure if that is contamination.
i saw no other colours and the smell was shroomy.
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gone-pear-shaped
Stranger than fiction
Registered: 10/30/17
Posts: 822
Last seen: 8 months, 2 days
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Re: Recognizing and dealing with contamination [Re: laruz]
#27385615 - 07/13/21 09:19 AM (2 years, 8 months ago) |
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Yikes! That needs to have some sections taken from it and transferred to grow out on other plates! That is so overgrown that we can't tell which parts are clean.
If you used that directly to colonize a jar, it may or may not do well. Do you have pictures of the jar?
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Enjoil
Registered: 10/29/20
Posts: 2,472
Last seen: 13 hours, 32 minutes
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That is his jar.
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Smartattack
C'mon man
Registered: 12/21/18
Posts: 3,779
Loc: A thought
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Re: Recognizing and dealing with contamination [Re: Enjoil]
#27385647 - 07/13/21 09:43 AM (2 years, 8 months ago) |
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--------------------
* Smarts videos
* Planet of the APES
I'm a fungal white supremacist.
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Enjoil
Registered: 10/29/20
Posts: 2,472
Last seen: 13 hours, 32 minutes
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Re: Recognizing and dealing with contamination [Re: laruz]
#27385648 - 07/13/21 09:43 AM (2 years, 8 months ago) |
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Looks fine to me… you should make you a few shoeboxes.
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gone-pear-shaped
Stranger than fiction
Registered: 10/30/17
Posts: 822
Last seen: 8 months, 2 days
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Re: Recognizing and dealing with contamination [Re: Enjoil]
#27385759 - 07/13/21 11:07 AM (2 years, 8 months ago) |
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Haha, I thought that was a glass petri dish! I've never seen jar photos of the top like that. It would not occur to me to remove the lid to check on its progress 
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