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Raven44
Entry not permitted to muggles



Registered: 12/07/13
Posts: 1,970
Loc: My sovereign reality bubble
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PDA few beginner questions
#21327334 - 02/25/15 10:29 AM (9 years, 6 days ago) |
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Recently been making my own agar and over coming all agar work nuisances that a beginner might encounter lol.. First question is, I made some PDYA and when I made it I used 75g potatoes simmered for 1hr with 250ml distilled water. That's equivalent to 300g potatoes and 1L distilled water. Then I read that I would want to use 300g potatoes to 2-3L water... Would anyone here consider these dishes I prepared to be more susceptible to contams by any means due to the fact that the potatoe water I prepared is more concentrated than recommended? I'm concerned cause I have no microscope to ensure my cultures are clean. Also, when these dishes germinated the PDA changes color in the area where spores germinated. Turning the PDA a darker tan color.. Its my first time w PDA, is this normal? Any input appreciated thank u much love
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PsyCLown89
Shroomaloomed



Registered: 08/18/14
Posts: 789
Loc: South Africa
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Re: PDA few beginner questions [Re: Raven44]
#21327392 - 02/25/15 10:40 AM (9 years, 6 days ago) |
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Post pictures so we can see exactly what it looks like, although often the agar will change colour (often goes lighter) when the nutrients are consumed by the myc.
I use instant mash potato powder and a drop of money in my PDA, I have found 1.25g potato powder, 1.25g agar and 200ml of water (normal tap) works quite well for germinating spores on. When doing transfers it might be best to make it a bit harder so a bit more agar.
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Raven44
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Registered: 12/07/13
Posts: 1,970
Loc: My sovereign reality bubble
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Re: PDA few beginner questions [Re: PsyCLown89]
#21327506 - 02/25/15 11:04 AM (9 years, 6 days ago) |
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My thought is, since the broth is more concentrated it has more nutes for contams possibly..? This info is very pertaining to my post thank u psyclown. This has been a prob for me, germinating dehydrated spores printed on paper from pikes place.. Great genentics(guy goes to Thailand, Brazil, ECT and collects wild specimens I talked to them about it..). So I've been experimenting w diff agar recipes to recently find out less water is good in recipe for germination. Great info thank u. I can't post pics for some reason. No computer on my phone.. When I go to click browse button it does nothing.. Can't browse to upload my photos.. Any input there..? Lol can I donate a buck and become a supporter to get an address I can send a text of the pic to lol? Not very computer/internet hip here.. I'd love to get pics up. Pretty sure I have mycelium growing, I've only got germination where I streaked spores, or where I streaked spores and put a drop of water on some of the dishes. The darker color to the PDA is only where germination has occurred and u can see growth. The darker regions reach out 5mm or so out all around from where the growth is. In other words, it seems the darker discoloration is from the mycelial growth possibly. Maybe from secretions advancing ahead of the mycelium.. I guess I'm askin if maybe anyone has done the same(made PDA broth more concentrated than recommended) and obtained a culture that was clean and/or didn't seem to have higher contam ratios attributed to the fact that the PDA broth was more concentrated than recommended.. Thanks for the input much love. In a tough spot here, been leaning/researching hands on past year, made many sacrifices to keep going, ran into contam probs do to improper preparation of LC, restarted everything, finances limited and dwindling, decisions critical at this point. I have MEA Petris that will germ prob in few days that I could use that are sure to be free of any concern. But they're 4 days behind these PDA dishes. If its not risky id like to use the PDA dishes now to expand, instead of wait on my MEA. In the end I might just expand both cultures but would rather not to save resources Petris and grain and such...
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Raven44
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Registered: 12/07/13
Posts: 1,970
Loc: My sovereign reality bubble
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Re: PDA few beginner questions [Re: Raven44]
#21327516 - 02/25/15 11:06 AM (9 years, 6 days ago) |
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Also, since I have no microscope I'm considering using a H2o2 agar recipe. Any input there? I'm doing all my work in front of a flow hood. And I'm doing agar work w out a microscope cause I'm aware it the best way to obtain a clean culture...
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PNWMusicMaker
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Registered: 10/18/14
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Loc: PNW
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Re: PDA few beginner questions [Re: Raven44]
#21327743 - 02/25/15 12:17 PM (9 years, 6 days ago) |
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Personally I think you're overcomplicating things. Check the link in my SIG, agar should be simple in my opinion.
-------------------- Live to change, change to live.
  
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Raven44
Entry not permitted to muggles



Registered: 12/07/13
Posts: 1,970
Loc: My sovereign reality bubble
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Raven44
Entry not permitted to muggles



Registered: 12/07/13
Posts: 1,970
Loc: My sovereign reality bubble
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Re: PDA few beginner questions [Re: Raven44]
#21327798 - 02/25/15 12:29 PM (9 years, 6 days ago) |
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If I had access to a microscope, id have zero need to ask what I've asked here. However I dont have a microscope or access to one. So I'm posting this realizing some won't be able to follow me here in my thought process, that's ok I understand but I'm posting in hopes that someone w the knowledge or first hand experience can give me advice. If not, I may just continue somewhat blindly hoping I'm not wasting resources... And in the future if I come across someone asking what I've asked here ill be able to give them advice. Thank u to any kind hearted open individuals who share their knowledge. Much love.
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PsyCLown89
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Registered: 08/18/14
Posts: 789
Loc: South Africa
Last seen: 2 years, 2 months
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Re: PDA few beginner questions [Re: Raven44]
#21327983 - 02/25/15 01:04 PM (9 years, 6 days ago) |
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Majority of us here do not use a microscope when working with agar, you even have a flowhood where majority make use of a SAB.
I am pretty sure you will be alright, agar really isn't that hard and no need to worry about it as much as you are. Take some instant potato powder, some agar powder and some water, mix it together in a pot and then sterilize it and pour it into some petri dishes or place it into a PP5 container and then sterilize that.
Simple really and it will work great, you will notice just about any contam that would be detrimental to your mushroom growing before using the culture on the agar to inoculate some substrate. 
If you are worried about resources and costs when it comes to agar, then I really think you are over complicating things. I am sure that using the more concentrated potato agar will be perfect. Since you are working with agar, you will most likely notice the contams before making use of it, however you could take this opportunity to try and isolate further and then do another transfer later on when you have made some new agar and petri dishes. Agar really isn't that expensive and will save you a lot of money in the long run, rather throw away a contaminated agar plate than a whole contaminated monotub as you did not check how clean your source (culture / spores) were on agar before hand. 
If it were me, I would use those potato agar petri dishes. Worst case scenario, they do contam and you learn from it and you can come back and post what happened here so anyone else wondering can find this thread and learn something too.
-------------------- My Trade List - Click ME!! [Updated: 02/05/2015]
"It's all in your head" "Haste makes waste!"
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Raven44
Entry not permitted to muggles



Registered: 12/07/13
Posts: 1,970
Loc: My sovereign reality bubble
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Re: PDA few beginner questions [Re: PsyCLown89]
#21328194 - 02/25/15 01:37 PM (9 years, 6 days ago) |
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Well thanks for ur input, its appreciated. I had already decided to just go for it w these PDYA dishes.. I realize agar is cheap, I'm just broke to the point where I can't afford any more Petris anytime soon. Maybe can't afford rent.my financial situation is where a lot of my concern comes from. I'm also under the impression that w agar sometime u CANNOT see contams until after u transfer agar wedges to a grain jar... I realize I'm.kinda asking a pointless question, I know. I'm just looking for educated input since its such a crucial time for me. Ty still tho..
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Raven44
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Registered: 12/07/13
Posts: 1,970
Loc: My sovereign reality bubble
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Re: PDA few beginner questions [Re: Raven44]
#21328234 - 02/25/15 01:44 PM (9 years, 6 days ago) |
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So ur saying in ur experience, when transferring agar to grain.. If your Petri appeared free of contam when u inoculated grain jars w agar wedges.. That those jars were always healthy cultures? Once ur jars are fully colonized, do u shake them to see if they re-colonize within 12-24 hrs. And if they DON'T re-colonize within 12-24 hrs they're considered contaminated? That's Paul stamets trick that works very well I've realized in my experience. Jars that DON'T re-colonize within 12-24 are very noticeable, have a foul smell if ur silly enough to smell them after opening them as I have before.. (I recommend shaking the jar and smelling it w the lid and filter still on..) oh and they never fully colonized again in my experience. The bad ones never fully re-colonize.
Edited by Raven44 (12/04/15 10:10 AM)
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Raven44
Entry not permitted to muggles



Registered: 12/07/13
Posts: 1,970
Loc: My sovereign reality bubble
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Re: PDA few beginner questions [Re: Raven44]
#22612322 - 12/04/15 10:11 AM (8 years, 2 months ago) |
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Ha ha old posts can b funny.
There was nothing wrong w my PDA.
Not too much to learn here sry lol.
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cronicr


Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 1 month
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Re: PDA few beginner questions [Re: Raven44]
#22612367 - 12/04/15 10:25 AM (8 years, 2 months ago) |
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--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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