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OfflinePSILOxPSYCHO
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Registered: 10/02/16
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Last seen: 7 years, 1 month
help with liquid culture
    #23715462 - 10/07/16 07:44 AM (7 years, 3 months ago)

so i realize the only way to see if a lc is clean is to knock something up. if i knocked brf cake jars up and they did good, would this mean my lc should be good to use for wbs, popcorn and rye? or is it still a crap shoot. ive herd brf cakes are pretty contamination resistant but does that matter? any help would be greatly appreciated it.


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Invisibleamidogen
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Registered: 05/07/16
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Re: help with liquid culture [Re: PSILOxPSYCHO]
    #23715604 - 10/07/16 08:39 AM (7 years, 3 months ago)

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The biggest trip of my life was realizing all of the events and actions described in posts made by this account were never real and had never actually happened, but were instead the delusional ramblings of a severely mentally ill human being. I just had to move on for my own good. I love you all.


Edited by amidogen (06/21/18 11:39 AM)


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OfflinePSILOxPSYCHO
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Re: help with liquid culture [Re: amidogen]
    #23715709 - 10/07/16 09:18 AM (7 years, 3 months ago)

Quote:

amidogen said:
It needs to be tested on agar. Period.




thats not true though....i dont think..... if i test it on grain it should show contamination as well, if there is any. am i not correct?....please correct me if im wrong. i realize agar is the best way to tell for contamination. i know this. thats not what im asking. my Question is. if my brf jars were inoculated with liquid culture and its 100% colonized with no contamination. does this mean i have a clean culture? if not please explain why.


Edited by PSILOxPSYCHO (10/07/16 09:54 AM)


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Invisibleamidogen
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Registered: 05/07/16
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Re: help with liquid culture [Re: PSILOxPSYCHO]
    #23715903 - 10/07/16 10:32 AM (7 years, 3 months ago)

.


--------------------
The biggest trip of my life was realizing all of the events and actions described in posts made by this account were never real and had never actually happened, but were instead the delusional ramblings of a severely mentally ill human being. I just had to move on for my own good. I love you all.


Edited by amidogen (04/19/18 11:06 AM)


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OfflinePSILOxPSYCHO
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Re: help with liquid culture [Re: amidogen]
    #23715994 - 10/07/16 11:03 AM (7 years, 3 months ago)

Quote:

amidogen said:
Well sure, I suppose you could test it on grains if you want. They'd probably be roughly as unforgiving as agar.

The reason your BRF cakes don't mean much is because BRF cakes can be inoculated with bacterial spawn and still produce fruits without turning green. Grains on the other hand aren't as forgiving. If you're looking to use your LC for anything besides PF cakes, you need to test it on agar. I don't see why you'd test it on a jar of grains. If it's a bad LC, you've just wasted a jar of grains. If you do it on agar, all you lose is a single plate.

Here's a better question: how did you make your LC? If you went straight from spore solution, it's practically guaranteed to be bacterial and will end up in your grains turning green. The only way to make LC without risking wasting it and everything you inoculate it with is to make it from a clean culture on agar. So which did you do, MS or agar?




A jar of grains means nothing to me. pennies really.

let me start this by saying everything was started from ms. you have to start somewhere right?
ive just started to grow these wonderful fruits so ms is all i have to start with.

that being said ms was used to create a few lcs, one of which i used to noc up a brf jar as well as two grain jars. My brf is about a day or so away from being 100% with no contamination, thats why my question about the brf. Which i kind of already knew the answer to.

As far as agar goes, i have started working with it.
I used a couple drops of lc to the agar and have a bit of growth, but I didnt have access to petri dishes for a month. still dont have em. so im using what i think they call 1/4 pint mason jars. tiny little guys. problem with that is, its hard to see what the growth is. it looks white and fuzzy, in patches(mycelium was injected onto surface by means of syring insides SAB.)

ive tried looking for pictures of patches of mycelium growing with no success. The growth is only small and hard to get any pictures of through the glass jars


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InvisibleMad Season
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Re: help with liquid culture [Re: PSILOxPSYCHO]
    #23716114 - 10/07/16 11:47 AM (7 years, 3 months ago)



This is what bacteria looks like on agar. Nice milky looking yellow blob, sometimes they're different colors too.



Another dish with several types of bacteria.



This is what a grain jar covered in bacteria looks like. Always test LCs on agar, if you want definitive results. Grains are definitely not the way to do it. Besides agar is fuckin easier than the pf tek.


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OfflinePSILOxPSYCHO
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Re: help with liquid culture [Re: Mad Season]
    #23716157 - 10/07/16 11:59 AM (7 years, 3 months ago)

you sir are my hero. thank you. ill get some pics of one of my agar dishes this evening when i do some more inoculations to agar. i can use my SAB and get a picture of the growth i have in one of my jars. maybe you can tell me if its fine or not. i use a patato flakes agar tek and it seems to work. id just like confirmation that it looks okay. you seem to answer me pretty quickly and i appreciate it. Thanks:grin: im pretty sure im doing okay as i did read for a couple months before i tried anything. i had everything but the petri dishes to start, so i may have jumped a bit fast but im learning.


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OfflinePSILOxPSYCHO
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Re: help with liquid culture [Re: PSILOxPSYCHO]
    #23721177 - 10/09/16 03:11 AM (7 years, 3 months ago)

Quote:

Mad Season said:


This is what bacteria looks like on agar. Nice milky looking yellow blob, sometimes they're different colors too.



Another dish with several types of bacteria.



This is what a grain jar covered in bacteria looks like. Always test LCs on agar, if you want definitive results. Grains are definitely not the way to do it. Besides agar is fuckin easier than the pf tek.




alright took me a couple days as I was busy but at last pictures of my liquid culture to agar growth. This was inoculated with a lc syring inside a SAB. The Inoculation was done through in a 1/4 pint mason jar which had a hole in the lid and tyvek under the the lid. My jars are also poured and then sterilized which is why the weird slant on the agar. I think the growth looks good but could you let me know what you think.


As well here is a picture of my popcorn that I Inoculated with the same liquid culture as well I think its got the same white fluffy/rhizo mycelium growth as my agar jars. so did i get lucky and get a clean culture my first try? i did follow all the teks to a T. have the teks written out in a book just so i didnt have to refer back to the computer to find what i was looking for. It was right there on paper in front of me.


So a couple questions. how does the agar and popcorn look. are they both good like i think.

how long should i wait to transfer to my next petri dish, should i let it grow out and start to transfer sectors to start isolation or should i transfer the known good piece to a clean petri dish and start isolation after that?


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Invisibleamidogen
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Re: help with liquid culture [Re: PSILOxPSYCHO]
    #23721418 - 10/09/16 08:15 AM (7 years, 3 months ago)

.


--------------------
The biggest trip of my life was realizing all of the events and actions described in posts made by this account were never real and had never actually happened, but were instead the delusional ramblings of a severely mentally ill human being. I just had to move on for my own good. I love you all.


Edited by amidogen (04/19/18 11:06 AM)


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OfflinePSILOxPSYCHO
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Re: help with liquid culture [Re: amidogen]
    #23721882 - 10/09/16 11:47 AM (7 years, 3 months ago)

You didnt answer any of my questions.....thanks, i guess any bit helps.....but i read a lot and decided popcorn was for me. Its cheap, Its readily available, and there is lots of teks explaining how to do a good popcorn tek. Im pretty good at following instrustion and so far thats proven to be so with what i think is all healthy mycelium growth. I am how ever looking for a more experienced opinions. so again   

How does everything look?

should i transfer now while I know its a clean culture and then start to sector for isolating sub-strains or should i wait till it grows out before starting my isolates. Thanks for the responses.


Edited by PSILOxPSYCHO (10/09/16 04:08 PM)


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InvisibleMad Season
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Re: help with liquid culture [Re: PSILOxPSYCHO]
    #23727572 - 10/11/16 07:49 AM (7 years, 3 months ago)

First pic looks like it has hella bacteria/yeast. Either that or its just a lot of sediment on the bottom. The white growths are all mycelium that's healthy tho. Transfer it and see what's up


--------------------
contam and car window art
How to shroomery like a pro! (Seriously, everyone read this!)
Improve your sterile techniques! (A comprehensive guide to agar)
Links upon links of literally EVERYTHING UP TO DATE

AMU Q&A
No trees were killed in the sending of this message. However, a large number of electrons were terribly inconvenienced.


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OfflinePSILOxPSYCHO
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Registered: 10/02/16
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Re: help with liquid culture [Re: Mad Season]
    #23728572 - 10/11/16 03:06 PM (7 years, 3 months ago)

Quote:

Mad Season said:
First pic looks like it has hella bacteria/yeast. Either that or its just a lot of sediment on the bottom. The white growths are all mycelium that's healthy tho. Transfer it and see what's up




Thanks for the reply Mad, i do think its sediment because i use potato flakes. great to know i got healthy mycelium :smile: i guess i got lucky my first try because this dish was started from my first ever liquid culture of GT. im stoked. ive done a bunch of reading and ordered some petri dishes and culture slants from ebay. 20pk of petri dishes(disposable) and 25 pack of slants(glass) for 50 taxes in delivered in a week. i did my first transfers last night, 3 from the most rizo spots in the mycelium.

if im correct i would keep transfering different areas or sectors until i see more of a mono culture and then test all the plates that have gotten to that stage to see which one is the best fruiting am i correct?
ive done lots of reading but no direct help except asking for help if i had contams or not. so far so good so good i think lol?


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OfflineLa Flama Blanca
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Re: help with liquid culture [Re: PSILOxPSYCHO]
    #23728711 - 10/11/16 04:06 PM (7 years, 3 months ago)

the slant tubes probably won't be of much use to you yet. people use those for really good/isolated cultures once they've found them, the search takes quite a while. i've heard people say it will take hundreds if not thousands of transfers to find a monoculture.

also, those 20 petris will go very, very fast. i use that many in one session typically.

also.... hard to tell from the pic, but everything looks good so far (though if i were you i'd avoid the growths that look very cottony/non-uniform. they're probably fine but could be mold). keep at it, you'll be amazed at how quickly agar work becomes nearly effortless. its the most important tool in this hobby though, so persist! :thumbup:


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