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blkbst
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Are all strains white? Is this contaminated? Help (pics)
#24904034 - 01/10/18 09:02 PM (7 years, 7 days ago) |
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First time cultivating, made agar with honey and potato flakes. Poured 3 Petri dishes (i'm using plastic ones). 2 dishes look somewhat similar, and one is a total mess, but looking at the photos here on the forums the mycelium doesn't look at all like that. At the same time this green thing is present on the 3 dishes, so i guess that's what i got from the syringe. Any ideas what to expect here? Is this green thing contaminated or is it what i got on the syringe i bought?


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Just_A_Noob



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Re: Are all strains white? Is this contaminated? Help (pics) [Re: blkbst]
#24904040 - 01/10/18 09:04 PM (7 years, 7 days ago) |
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Looks like there is some myc in the second one but yeah all those are contaminated. Did you use a SAB?
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mushpunx
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: Just_A_Noob]
#24904092 - 01/10/18 09:19 PM (7 years, 7 days ago) |
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That is absolutely mold.
Yea, describe your process a bit for us please
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Tormato  
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: mushpunx]
#24904099 - 01/10/18 09:22 PM (7 years, 7 days ago) |
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UGH! Trich and Pin mold
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Philosopherski
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: Tormato]
#24904106 - 01/10/18 09:24 PM (7 years, 7 days ago) |
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Sorry mate
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Subnet Mask


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Re: Are all strains white? Is this contaminated? Help (pics) [Re: Philosopherski]
#24904108 - 01/10/18 09:26 PM (7 years, 7 days ago) |
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Impressive.
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blkbst
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: mushpunx]
#24904129 - 01/10/18 09:34 PM (7 years, 7 days ago) |
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I PCed the agar, poured it under a SAB, let it cool, and gave a drop or two from a spore vial i got from a store I didn't expect all 3 dishes to fail so badly i'll be extra careful next try thanks for the replies
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Chief_Slapaho
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: blkbst]
#24904261 - 01/10/18 10:20 PM (7 years, 7 days ago) |
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Honestly, it's probably the spore solution that carried the mold. Assuming your technique was good. You can pretty much always assume that spore prints and thus spore solutions have some contaminates.
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UnsungHero
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: Chief_Slapaho]
#24904272 - 01/10/18 10:26 PM (7 years, 7 days ago) |
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Check your sterile technique, step up your SAB game and give it another go! Don't give up it's part of the process.
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ShiVersblood
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: blkbst]
#24904371 - 01/10/18 11:09 PM (7 years, 7 days ago) |
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It looks blue
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DavieDonna
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: ShiVersblood] 1
#24904717 - 01/11/18 02:15 AM (7 years, 6 days ago) |
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That's some hardcore trich. Makes me angry just looking at it.
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LemurLemur
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: blkbst]
#24904747 - 01/11/18 03:02 AM (7 years, 6 days ago) |
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In that first one is that were u put your spore solution?? If so its the syringe, try a print i find it much much much better than spore solution, i think syringes are for making cakes.
Infact try cakes or grains through SHIPS to get some mycilium then put a sample to agar and tranfer away.
Get a close up of that second 1 with the flash from underneath and on top, with so much tams u wanna be sure.
Spray the inside of your sab with soapy water with fine mist wait for it to settle and slowly move your hands u should move as fast as a chameleon, dont open your dishes more than u need n only open the dish sleeve in the sab, just speeze them in through a arm hole like cookie doe from 1 end. Look up Bod's n c10's agar guides.
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Boxer
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: LemurLemur]
#24904777 - 01/11/18 04:29 AM (7 years, 6 days ago) |
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The first plate looks like penicillium, the 2nd has trich & pin mold and that's mycelium on the left. The third has trich and the white blotches are bacterial. Based on how wide spread the contamination is I would say it's you're syringe that's contaminated. You won't get anything clean from it more than likely. if getting another print/syringe isn't an option, I would transfer from the mycelium on the 2nd plate and see if you can clean it up via several transfers.
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blkbst
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: Boxer]
#24905947 - 01/11/18 04:44 PM (7 years, 6 days ago) |
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I'm doing another pour right now, waiting for the agar to cool. I'll try getting a piece of the "mycelium" in the second one and put it in 2 new dishes, and try again with the syringe to see if it really is contaminated I'll update next week after this new pour grows
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Cakewonder
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: blkbst]
#24906048 - 01/11/18 05:36 PM (7 years, 6 days ago) |
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Quote:
blkbst said: I PCed the agar, poured it under a SAB, let it cool, and gave a drop or two from a spore vial i got from a store I didn't expect all 3 dishes to fail so badly i'll be extra careful next try thanks for the replies 
He used a SAB
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Edited by Cakewonder (01/11/18 10:47 PM)
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mushpunx
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: Cakewonder]
#24906070 - 01/11/18 05:47 PM (7 years, 6 days ago) |
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Quote:
Cakewonder said:
Quote:
blkbst said: I PCed the agar, poured it under a SAB, let it cool, and gave a drop or two from a spore vial i got from a store I didn't expect all 3 dishes to fail so badly i'll be extra careful next try thanks for the replies 
It is close to impossible to do pour in open air. Use the NOPOUR TEK.
? He said he poured it in a still air box, not open air. You can have the same success rate pouring in a SAB as in front of a flow hood. I learned in a SAB.
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streamer

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Re: Are all strains white? Is this contaminated? Help (pics) [Re: mushpunx]
#24906177 - 01/11/18 06:43 PM (7 years, 6 days ago) |
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I'd also pour an extra plate to see if the contams are coming from 'the pour' itself. A single drop of MS in the center is all that's needed. Your contams seem very widespread. Good luck!
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SimpleFarmer



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Re: Are all strains white? Is this contaminated? Help (pics) [Re: streamer]
#24906326 - 01/11/18 07:41 PM (7 years, 6 days ago) |
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How long are you pcing?
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Cakewonder
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: SimpleFarmer]
#24906689 - 01/11/18 10:34 PM (7 years, 6 days ago) |
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I should sau sorry, I am no agar expert, there are other people who know this better and I should leave it to them to help him. I am sorry.
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Edited by Cakewonder (01/11/18 11:00 PM)
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blkbst
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: SimpleFarmer]
#24907297 - 01/12/18 07:55 AM (7 years, 5 days ago) |
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Quote:
SimpleFarmer said: How long are you pcing?
45~60 mins. My PC doesn't make it to 15 psi, it only goes up to ~11 and I'm a bit reluctant to tape some coins on the valve trying to get higher pressure.
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mushpunx
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: blkbst]
#24907307 - 01/12/18 08:04 AM (7 years, 5 days ago) |
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Are you wrapping the edge of your plates after inoculation?
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blkbst
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: mushpunx]
#24907338 - 01/12/18 08:19 AM (7 years, 5 days ago) |
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The first time i totally forgot about it, and once out of the SAB i noticed, so i just leave them that way, didn't want to make things worse trying to seal them in an unclean space or manipulate the dishes more than what's necessary. But this time i did seal them with tape while under the SAB, hope this ones do better
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mushpunx
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: blkbst]
#24907378 - 01/12/18 08:34 AM (7 years, 5 days ago) |
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Don't use tape. Wrap the edge tight with Parafilm or plastic cling wrap (saw 1" off the end of a roll). Uninoculated plates should be kept in their sleeve or in sterile ziplocks.

Here is a photo of a plate wrapped with laboratory Parafilm and one wrapped with cling wrap
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blkbst
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: mushpunx]
#24907645 - 01/12/18 10:39 AM (7 years, 5 days ago) |
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Nice idea about cutting a piece of a cling wrap roll. I'll definitely do that the next time. Just hope that the tape will at least help this time
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blkbst
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: blkbst]
#24910728 - 01/13/18 12:00 PM (7 years, 4 days ago) |
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(Sorry for the bump) It looks like i managed to isolate whatever white stuff was growing on the dish from the second pic, it grew really fast, it has white hair thingies, not sure if this is what i'm supposed to get. The big problem now is that my dishes got fogged, and i can't get a good picture. In fact i can't see in detail what's going on inside, is there anything i can do to defog the inside of the dish? or i'm doomed to open and remove fog by hand? i did my best trying to get a picture that shows anything, i played with curves and managed to make the "mycelium" more visible. Not even sure if it's mycelium 
hope someone can help guys 

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Skizor1337
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Re: Are all strains white? Is this contaminated? Help (pics) [Re: blkbst]
#24910832 - 01/13/18 12:45 PM (7 years, 4 days ago) |
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Quote:
blkbst said:
Quote:
SimpleFarmer said: How long are you pcing?
45~60 mins. My PC doesn't make it to 15 psi, it only goes up to ~11 and I'm a bit reluctant to tape some coins on the valve trying to get higher pressure.
What pressure cooker are you using?
I use a Presto cooker that doesn't even have a PSI meter on it and it does just fine bro.
It's a 6 quart stainless and i can PC my media bottle
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