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byo_shroom
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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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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




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did you spray your sab with soapy water prior to use?
-------------------- Song Of Healing
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The Chinese word for nature is zìrán and it means that of which is of itself.

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mushboy
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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
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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
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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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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
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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
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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
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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



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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
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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
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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
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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



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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
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Keeps contaminants from getting into the plate Holds them together nicely too
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Shrooms4life
TrippyShaman



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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
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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
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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
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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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mushboy
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Re: Contaminated agar? [Re: mushpunx]
#24917655 - 01/16/18 07:37 AM (7 years, 1 day ago) |
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for sho! wrap occupied media asap!
my point was more for pour newbies. if you jam your blank media inside a ziplock or whatever.. and it molds?? the brain can fire off a million reasons besides the real issue.(your pouring sucked) but your brain can rationalize it..
'nahh.. my pouring is legit, these molded when i put them in the ziplocks.. i must of banged em around to much'
for me? that kinda shit breeds noob mentality all day in my head so i gotta be on my own top.
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mushpunx
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Re: Contaminated agar? [Re: mushboy]
#24917662 - 01/16/18 07:44 AM (7 years, 1 day ago) |
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Haha yea I hear you man!
Pouring is so easy. I think I lost one plate my first pour but never again. I love to pour!
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mushboy
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Re: Contaminated agar? [Re: mushpunx]
#24917672 - 01/16/18 07:51 AM (7 years, 1 day ago) |
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my first pour was 90% contam. bod quipped 'wtf did you pour in open air lolwut?' ... 
but after that first botched pour, the second time was ok.. by the 3rd... i never..ever ever contam from pouring.
out of 1000s
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Shrooms4life
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Re: Contaminated agar? [Re: mushboy]
#24917752 - 01/16/18 08:47 AM (7 years, 1 day ago) |
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Alright thanks for the info guys, I'll get some Cling wrap today after work and make some new agar for transfers and wrap them.
Do you guys wrap them in the sab? Or once all the work is done?
Also any ideas on how to cut back on so much condensation in the top of my petris when I pour? Yours look killer mushboy!
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Mitchell1992
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Need to let the agar cool more before you pour it to avoid condensation can’t rember the temp but I generally hold it to the top of my sab in the bottle and if it don’t leave any condensation I pour it.
I wrap them as soon as I take them out with no issues
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mushboy
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Quote:
Shrooms4life said: Also any ideas on how to cut back on so much condensation in the top of my petris when I pour? Yours look killer mushboy!
stack em as they cool. nd what he said about proper pouring temps. they shouldnt really condense at all. people who drink coffee say to put the hot mug on top of a foggy dish or some shit i dunno.
or you dont give fuck cause its just a few petris anyway?
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Shrooms4life
TrippyShaman



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Re: Contaminated agar? [Re: mushboy]
#24918060 - 01/16/18 11:17 AM (7 years, 1 day ago) |
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Quote:
mushboy said:
Quote:
Shrooms4life said: Also any ideas on how to cut back on so much condensation in the top of my petris when I pour? Yours look killer mushboy!
stack em as they cool. nd what he said about proper pouring temps. they shouldnt really condense at all. people who drink coffee say to put the hot mug on top of a foggy dish or some shit i dunno.
or you dont give fuck cause its just a few petris anyway?
Well I typically wouldn't give 2 fucks but some of the plates are almost impossible to see into or take pics of, and some others have had a drop or 2 fall into the agar itself.
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Mitchell1992
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Don’t think it matters much mate isn’t going to do much harm
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Shrooms4life
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I'm just saying id rather be able to pour without blocking my view of the culture lol. Like i said I'm new to the process these are my 1st plates so I'm going to need to take pics and run em past the pros first.
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byo_shroom
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Re: Contaminated agar? [Re: mushpunx]
#24919019 - 01/16/18 06:19 PM (7 years, 1 day ago) |
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Quote:
mushpunx said:
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.
With the grains I rinsed and then soaked for 24 hours. Then bought to the boil and simmered for 10 mins. Then strained, and added to my jars. Then steam sterilised 3 times for two hours over 3 days (it was actually more like a total of 7 hours over the 3 days).
I read that somewhere and others commented saying they had success will less. Ideally I'd have the PC, but I wanted to try this on just 3 jars and see how I go.
The jars were also agar to grain transfer from the first culture off the spoor print. I was just too eager to wait around. Fingers crossed they stay contamination free.
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mushpunx
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You definitely want to make at least one transfer after germination before putting to grain.
Are you saying you loaded your jars directly after straining? You are supposed to give them a shake after draining and let them steam dry, giving a shake occasionally.
Once you can put a handful on a paper towel and they don't leave a wetspot they are ready to be loaded into jars.
If you are using grain like rye and do t let them steam dry after boiling the jars will be way too wet and prone to bacterial problems
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byo_shroom
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Re: Contaminated agar? [Re: mushpunx]
#24919723 - 01/17/18 12:16 AM (7 years, 23 hours ago) |
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Yes, they steam dried. Like you said, I put some on a paper towel to test if they left a mark behind.
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mushpunx
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Well then you're prep should be good!
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