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OfflineToobaked
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First time agar
    #26970883 - 10/05/20 04:27 PM (3 years, 3 months ago)

I eagerly scraped a small print onto a couple of ore made petri’s. After the fact learning that it’s best to concentrate the sores i one area. Can I still salvage anything from these ?


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OfflineForresterM
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Re: First time agar [Re: Toobaked] * 1
    #26970915 - 10/05/20 04:44 PM (3 years, 3 months ago)

I would toss them.  If you're trying to clean up spores you want to take transfers of mycelium a lot earlier.  And like you probably know now, try to get as few, not as many, spores on there as possible.


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Have some medicinal mushrooms and want to get the most out of them?  Try this double extraction method.


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Offlineshroomshine
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Re: First time agar [Re: Forrester]
    #26970938 - 10/05/20 04:58 PM (3 years, 3 months ago)

why is there no lid on these?


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OfflineToobaked
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Re: First time agar [Re: shroomshine]
    #26970965 - 10/05/20 05:24 PM (3 years, 3 months ago)

I took it off to get a clear picture


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Offlineshroomshine
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Re: First time agar [Re: Toobaked] * 1
    #26970980 - 10/05/20 05:37 PM (3 years, 3 months ago)

I was wondering lol
if you ever have something transferable, don't take the lid off to take pictures. open air is pretty nasty.

when I started agar I made spore syringes, put a drop on a loop and made "Z" plates. I found that easier.

good luck! :smile:


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OfflineForresterM
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Re: First time agar [Re: shroomshine]
    #26971519 - 10/06/20 02:42 AM (3 years, 3 months ago)

Quote:

shroomshine said:
why is there no lid on these?




Oh boy... good catch.

OP not to sound rude but I think you should do a bit more reading on agar and sterile technique :smile:


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Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here.
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Have some medicinal mushrooms and want to get the most out of them?  Try this double extraction method.


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InvisibleMateja
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Re: First time agar [Re: Toobaked]
    #26971557 - 10/06/20 03:59 AM (3 years, 3 months ago)

Quote:

Toobaked said:
Can I still salvage anything from these?



I don't see any Cubensis mycelium on those plates, only bacteria and mold.
You should probably read up on how to properly work with agar plates inside a SAB if you want to get things to work out for you. Gl


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InvisibleJHOVA
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Re: First time agar [Re: Mateja]
    #26971839 - 10/06/20 10:13 AM (3 years, 3 months ago)

Read these threads and do what c10 says. You only meed a single drop of spore solution not a whole cc of liquid.

:dogpipe:



https://www.shroomery.org/forums/showflat.php/Number/21922023

https://www.shroomery.org/forums/showflat.php/Number/18430998

Quote:



Working with Spores:

since we fruit in open air, we can assume that spores are dirty. if someone sends you a spore print, you can assume it is going to have contams on it in addition to the spores you want to cultivate. this does not mean you got a bad print, it means you got a NORMAL print. the same thing applies to syringes. for this reason, when inoculating something with spores, it is a good idea to use as little as possible, since we only need a few spores and the more solution/spores we use, the more contams will be present as well. i recommend people never putting spores directly to grain, since it is so easy to culture the contams

a sterile jar of grain is basically the microbiological jackpot for anything that can digest it (fungi, bacteria, etc), so we need to take care to reduce the chances of culturing the wrong microorganisms. basically, ALWAYS germinate your spores first on agar before applying them to grain/lc/etc.

by using streaking techniques, you can easily clean up even the dirtiest of prints or spore syringes. you basically are using a loop to spread spores VERY thin across the surface of agar, creating various zones with different concentrations of spores. this is extremely useful for cleaning up a print, since you spread the contams thin while you spread the spores. this is also a great way to obtain isolates about 100 transfers sooner than you would get from a blob of spores, since the later zones have very few strains present in each colony. by doing one of these serial dilution style streaks, you end up with hundreds of colonies to choose from, of various genetic diversity, which gives you lots of chances to select a vigorous, organized culture

Quote:

In microbiology, streaking is a technique used to isolate a pure strain from a single species of microorganism, often bacteria(fungi in our case). Samples can then be taken from the resulting colonies and a microbiological culture can be grown on a new plate so that the organism can be identified, studied, or tested.

Streaking is rapid and ideally a simple process of isolation dilution. The technique is done by diluting a comparatively large concentration of bacteria to a smaller concentration. The decrease of bacteria should show that colonies are sufficiently spread apart to effect the separation of the different types of microbes. Streaking is done using a sterile tool, such as a cotton swab or commonly an inoculation loop. Aseptic techniques are used to maintain microbiological cultures and to prevent contamination of the growth medium.There are many different types of methods used to streak a plate. Picking a technique is a matter of individual preference and can also depend on how large the number of microbes the sample contains.

The three-phase streaking pattern, known as the T-Streak, is recommended for beginners.The streaking is done using a sterile tool, such as a cotton swab or commonly an inoculation loop. The inoculation loop is first sterilized by passing it through a flame. When the loop is cool, it is dipped into an inoculum such as a broth or patient specimen containing many species of bacteria. The inoculation loop is then dragged across the surface of the agar back and forth in a zigzag motion until approximately 30% of the plate has been covered. The loop then is re-sterilized and the plate is turned 90 degrees. Starting in the previously streaked section, the loop is dragged through it two to three times continuing the zigzag pattern. The procedure is then repeated once more being cautious to not touch the previously streaked sectors. Each time the loop gathers fewer and fewer bacteria until it gathers just single bacterial cells that can grow into a colony.The plate should show the heaviest growth in the first section. The second section will have less growth and a few isolated colonies, while the final section will have the least amount of growth and many isolated colonies.




examples of streaked plates. different labs use different standards for streaking. colonies thin as the dish rotates counter clockwise


the way i like to do it is very similar to what is described above. basically, take your loop (bod's DIY loop tek) and scrape off a small amount of spores from a print (or drip a drop from a syringe directly onto the loop) and streak a side of the plate like below. then flame the loop again, DONT SCRAPE MORE SPORES, rotate the dish, and drag the loop from the area you streaked previously to streak zone 2, then flame and repeat

this is the streaking technique i like to use. it is tremendously handy for cleaning up spore prints, isolation projects, and even cleaning up old LCs


if you are using a spore syringe as your starting point (or a LC you are trying to refresh), you want to be super careful not to add any additional liquid to the plate, otherwise it will roll around and distort the streaking pattern. so make sure you dont drip over the plate, you shouldnt be putting any more liquid into the plate than what sticks to the loop

Conclusion:


so that is pretty much it!! a bit lengthy, but i think i covered most of what i wanted to cover for my post #1000. please let me know if yall have any questions or need clarification about anything :smile:

i also welcome any additional tips & tricks, feedback, criticism, discussion, debate, haters (:kiss:), or critical thinking :wink:

thanks everyone for everything yall have done to make this place such an incredible resource. im so grateful to live in an age where there is an online community to collaborate with on our hobbies :smile:

warm regards,
c10




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OfflineNedRise
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Registered: 09/30/20
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Re: First time agar [Re: Toobaked]
    #26971883 - 10/06/20 10:39 AM (3 years, 3 months ago)

Quote:

Toobaked said:
I eagerly scraped a small print onto a couple of ore made petri’s. After the fact learning that it’s best to concentrate the sores i one area. Can I still salvage anything from these ?




what the fuck ... LMAO :grin:


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