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byo_shroom
Stranger

Registered: 04/29/17
Posts: 43
Last seen: 5 years, 4 months
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Contaminated agar?
#24917329 - 01/16/18 12:50 AM (7 years, 1 day ago) |
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Hi,
This is my first shroom grow. I transferred from a spore print to agar dishes. Some of which were in the mini mason jars, others were in sterile petri dishes.
Curiously, the mini jars grew far quicker than the agar dishes. Could this be due to the 1/4" gas exchange hole that was filled with poly fill?
Of the 5 petri dishes I'm working on, 3 were agar to agar transfer (2 of which maybe contaminated) and 2 were spore print to agar (one appears contaminated).
I used a SAB, is it common to get this high contamination rate with petri dishes? Both the glass jar agar grows seemed contamination free. I would have expected the reverse.
Pictures below:
1.

2.

3. This one I'm not sure quite what it is. It appeared from underneath the agar:

Also, I steam sterilised the rye berries. I read mixed opinions on steam sterilisation and I wanted to satisfy my own curiosity about its effectiveness (plus I don't have a PC yet, so I really didn't have a choice).
So far the grains look healthy. I transferred agar wedges to jars about 4 days ago. I've read a few comments saying to shake up the jars. At what point should I do that and what's the benefit of this?
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Mitchell1992
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Registered: 10/07/17
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Last seen: 2 years, 5 days
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You’ll always get contamination from spores that’s the reason for the agar to transfer away from it, make sure your hands are not over the dish when doing transfers or prints open the lid slightly and do your thing
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Ziran
The Hero of Time




Registered: 02/03/16
Posts: 6,034
Loc: Temple of Time
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did you spray your sab with soapy water prior to use?
-------------------- Song Of Healing
Updated Pf Tek Guide
Ziran's Teks
AMU Q&A Thread
The Chinese word for nature is zìrán and it means that of which is of itself.

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mushboy
modboy



Registered: 04/24/05
Posts: 35,378
Loc: eating the cats
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Re: Contaminated agar? [Re: Ziran] 1
#24917348 - 01/16/18 01:12 AM (7 years, 1 day ago) |
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Not bad for a first time. Heres my first time.
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Ziran
The Hero of Time




Registered: 02/03/16
Posts: 6,034
Loc: Temple of Time
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Re: Contaminated agar? [Re: mushboy]
#24917425 - 01/16/18 02:41 AM (7 years, 1 day ago) |
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Quote:
mushboy said: Not bad for a first time. Heres my first time.


lol. thats nuts. is pour agar that much harder?
-------------------- Song Of Healing
Updated Pf Tek Guide
Ziran's Teks
AMU Q&A Thread
The Chinese word for nature is zìrán and it means that of which is of itself.

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byo_shroom
Stranger

Registered: 04/29/17
Posts: 43
Last seen: 5 years, 4 months
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Re: Contaminated agar? [Re: Ziran]
#24917479 - 01/16/18 04:22 AM (7 years, 1 day ago) |
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Quote:
mushboy said: Not bad for a first time. Heres my first time.

Lol, suddenly feel much better, thanks!
Quote:
Ziran said: did you spray your sab with soapy water prior to use?
Yes, wiped it over with antibacterial wipes. Also used disposable latex gloves and used antibacterial gel on those too.
Although, I used an arts and crafts scalpel. It wasn't sterile out of the wrapper, but I used some rubbing alcohol on it and flame sterilised the blade.
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Mitchell1992
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Registered: 10/07/17
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Last seen: 2 years, 5 days
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They look alright mate tbh second picture down looks like you dropped some shit on doing the transfer! Is the third pic not just air under the agar? Can’t see on The phone
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byo_shroom
Stranger

Registered: 04/29/17
Posts: 43
Last seen: 5 years, 4 months
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Mitchell1992 , you could be right (trapped air). It's under the agar, not growing in the space between the agar and lid. I'll watch it over the next couple of days and see how it changes. Thanks.
I probably need to find some ideal looking cultures. I've been following teks from YouTube, it doesn't help that I don't have a point for comparison.
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Shrooms4life
TrippyShaman



Registered: 06/08/17
Posts: 1,376
Loc: Under the Viel
Last seen: 30 days, 15 hours
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So since I'm also new to agar I'll jump in here so we can all learn from each other, I currently have 9 plates going at different stages, I've done some that I poured myself in my shitty sab and I've also got a couple that are no pours in smaller then pint jars. I made my own self healing injection port lids and some sfd filters on them and they work like a charm!
I actually prefer the jars over the petris because maybe I'm just new to the process or pouring my agar too hot but my pours always seem to get lots of condensation in them mak ing it really hard to take pics or spot growth and contams. The other ones I made myself are crystal clear with 0 condensation at all because of the filter I think.
I currently have 2 plates of lgt from a print I got from a member here and some other random syringes to agar, then I've also got 4 plates that were 4th flush clone tissue to agar and they are taking off now, I'll take better pics when I get off work for you guys to critique.
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mushpunx
Fungus Punk



Registered: 04/20/14
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Are you wrapping the edge of the plates tightly with cling wrap or parafilm? Here's a photo of each, although the cling wrap one is sloppily wrapped. I saw a 1" off the end of a roll of cling wrap
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Shrooms4life
TrippyShaman



Registered: 06/08/17
Posts: 1,376
Loc: Under the Viel
Last seen: 30 days, 15 hours
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Re: Contaminated agar? [Re: mushpunx]
#24917596 - 01/16/18 06:44 AM (7 years, 1 day ago) |
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Quote:
mushpunx said: Are you wrapping the edge of the plates tightly with cling wrap or parafilm? Here's a photo of each, although the cling wrap one is sloppily wrapped. I saw a 1" off the end of a roll of cling wrap

If the question is pertaining to me then the answer is no i haven't been, I have been putting them in zip lock bags cause I saw a few others doing the same and didn't have cling or parafilm.
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mushboy
modboy



Registered: 04/24/05
Posts: 35,378
Loc: eating the cats
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Re: Contaminated agar? [Re: Ziran]
#24917599 - 01/16/18 06:46 AM (7 years, 1 day ago) |
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Quote:
Ziran said:
Quote:
mushboy said: Not bad for a first time. Heres my first time.


lol. thats nuts. is pour agar that much harder?
Actually its easy as fuck. But first times are always 'first times'
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mushpunx
Fungus Punk



Registered: 04/20/14
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Re: Contaminated agar? [Re: mushpunx]
#24917601 - 01/16/18 06:51 AM (7 years, 1 day ago) |
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Don't use wipes in your SAB. Just lightly spritz the walls and air with soapy water , let it settle about 10 minutes before using it. The air in the SAB will never be sterile, you just have to keep it still, and never move anything unsterile (like your gloves) over top open media, that's how it works. The soapy water just helps catch contams and help the air settle down faster.
If your spore print was dirty, you can get contamination on your germination plates yes, but the beauty of agar is that we can transfer away from it. I make at least 2 transfers before using a culture as inoculant.
Transfer as soon as you see clean myc, don't let it grow to touch any bacteria spots or mold colonies. If I see green mold on a plate I generally do not use those ones.
As far as your jars, how long did you steam sterilize them? Grains need at least 8 hours of steam if you aren't using a PC.
I only shake my grain jars after I have a colonized portion about the size of my closed fist. This redistributes the myc and the jars will colonize very fast afterwards.
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mushpunx
Fungus Punk



Registered: 04/20/14
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Quote:
Shrooms4life said:
Quote:
mushpunx said: Are you wrapping the edge of the plates tightly with cling wrap or parafilm? Here's a photo of each, although the cling wrap one is sloppily wrapped. I saw a 1" off the end of a roll of cling wrap

If the question is pertaining to me then the answer is no i haven't been, I have been putting them in zip lock bags cause I saw a few others doing the same and didn't have cling or parafilm.
No it was for the OP. But you need to wrap your plates.
Uninoculated plates can go back into their sleeve (so long as you keep the lids from opening), they can be wrapped or placed into sterile ziplock bags (I do 4 at a time. Fresh ziplocks are sterile).
But after inoculation the edge needs to be wrapped. Like I said saw a 1" section off the end of a roll of cling wrap and wrap the edge of the plates around a few times tightly. Or you can use labratory parafilm.
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Shrooms4life
TrippyShaman



Registered: 06/08/17
Posts: 1,376
Loc: Under the Viel
Last seen: 30 days, 15 hours
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Re: Contaminated agar? [Re: mushpunx]
#24917617 - 01/16/18 07:03 AM (7 years, 1 day ago) |
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Quote:
mushpunx said:
Quote:
Shrooms4life said:
Quote:
mushpunx said: Are you wrapping the edge of the plates tightly with cling wrap or parafilm? Here's a photo of each, although the cling wrap one is sloppily wrapped. I saw a 1" off the end of a roll of cling wrap

If the question is pertaining to me then the answer is no i haven't been, I have been putting them in zip lock bags cause I saw a few others doing the same and didn't have cling or parafilm.
No it was for the OP. But you need to wrap your plates.
Uninoculated plates can go back into their sleeve (so long as you keep the lids from opening), they can be wrapped or placed into sterile ziplock bags (I do 4 at a time. Fresh ziplocks are sterile).
But after inoculation the edge needs to be wrapped. Like I said saw a 1" section off the end of a roll of cling wrap and wrap the edge of the plates around a few times tightly. Or you can use labratory parafilm.
Gotcha, what's the purpose of wrapping the plates in cling or parafilm?
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mushpunx
Fungus Punk



Registered: 04/20/14
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Keeps contaminants from getting into the plate Holds them together nicely too
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Shrooms4life
TrippyShaman



Registered: 06/08/17
Posts: 1,376
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Last seen: 30 days, 15 hours
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Re: Contaminated agar? [Re: mushpunx]
#24917635 - 01/16/18 07:18 AM (7 years, 1 day ago) |
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Quote:
mushpunx said: Keeps contaminants from getting into the plate Holds them together nicely too
Gotcha thanks brother I'll have to get some Cling wrap then, does the ziplocks not do the same thing? Also I left 4 of my last clone tissue plates out in the sab not in bags just covered by the top of the petris should I cling em all up asap?
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mushboy
modboy



Registered: 04/24/05
Posts: 35,378
Loc: eating the cats
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i wouldnt wrap or ziplock blank media. just let it sit out in the SAB or in empty shoeboxes. they should not out right mold.
if they do you have a serious problem or your technique is jacked. but if you jammed them into ziplocks? its hard to pinpoint issues.. if you even get any.
 i pour and leave em alone. grab them as you need them. they will be fine inside your SAB... suppose if your SAB is mobile then you cant just leave em out.. soo.... do what you gotta do i guess.
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mushpunx
Fungus Punk



Registered: 04/20/14
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Last seen: 2 months, 7 days
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I would , yea. They should be okay in ziplocks but once inoculated it really is best to wrap them proper.
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mushpunx
Fungus Punk



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Re: Contaminated agar? [Re: mushboy]
#24917653 - 01/16/18 07:36 AM (7 years, 1 day ago) |
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Quote:
mushboy said: i wouldnt wrap or ziplock blank media. just let it sit out in the SAB or in empty shoeboxes. they should not out right mold.
if they do you have a serious problem or your technique is jacked. but if you jammed them into ziplocks? its hard to pinpoint issues.. if you even get any.
 i pour and leave em alone. grab them as you need them. they will be fine inside your SAB... suppose if your SAB is mobile then you cant just leave em out.. soo.... do what you gotta do i guess.
I think its best to put them back in the sleeve they came with. I don't go to the trouble of wrapping blank media though. Everyone stores their blank plates differently.
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