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Invisiblemaxmush
Always learning...
Registered: 06/13/20
Posts: 440
Why zig-zag spores to agar?
    #26843711 - 07/24/20 10:25 PM (3 years, 6 months ago)

I understand this is common practice and I have done myself, but what exactly is the rationale?


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OfflinePsilicon
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Re: Why zig-zag spores to agar? [Re: maxmush] * 1
    #26843734 - 07/24/20 10:49 PM (3 years, 6 months ago)

Streaking in a zigzag is used for isolation. The spores deposited onto the agar drop off logarithmically as long as you don’t pick up your loop, and it’s easier to zigzag than to do a perfect swirl without touching a place you’ve already touched. By the end of the zigzag you should see isolated colonies, which is the goal

I just put ‘em in the fucken middle though


--------------------
Agar - what, why and how?  Everything a beginner needs to know.
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Edited by Psilicon (07/24/20 10:55 PM)


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OfflineCamera93
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Re: Why zig-zag spores to agar? [Re: maxmush] * 2
    #26843738 - 07/24/20 10:51 PM (3 years, 6 months ago)

you create a clear path of where you streaked
so you know where to look for germination, if you get some satellite growth outside of the pattern, its likely not myc from the species you are attempting to cultivate

thats my reason

there is actually a method for streaking plates and getting isolated colonies:

Quote:

c10h12n2o said:
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


warm regards,
c10




you can read the whole write up here c10's Agar Guide + Tips & Tricks


--------------------
All I need are some tasty waves, a cool buzz, and I’m fine.

Whatever you decide won’t really impact our survival
Close your eyes, and do the best that you can



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Invisiblemaxmush
Always learning...
Registered: 06/13/20
Posts: 440
Re: Why zig-zag spores to agar? [Re: Psilicon]
    #26843741 - 07/24/20 10:52 PM (3 years, 6 months ago)

I am assuming the purpose is to have *less* spores at the end of the streak?


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Disclaimer: all information presented is intended for educational purposes only. All photos are only representations and not directly from the user.


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Invisiblemaxmush
Always learning...
Registered: 06/13/20
Posts: 440
Re: Why zig-zag spores to agar? [Re: maxmush]
    #26843744 - 07/24/20 10:53 PM (3 years, 6 months ago)

Thanks Camera93 helpful


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Disclaimer: all information presented is intended for educational purposes only. All photos are only representations and not directly from the user.


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OfflinePsilicon
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Registered: 08/26/12
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Re: Why zig-zag spores to agar? [Re: maxmush]
    #26843750 - 07/24/20 10:58 PM (3 years, 6 months ago)

Quote:

maxmush said:
I am assuming the purpose is to have *less* spores at the end of the streak?



Yes.


--------------------
Agar - what, why and how?  Everything a beginner needs to know.
Oat Prep Tek
Bored?  Please take one of my experiments off my hands.


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Invisiblec10h12n2o
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Re: Why zig-zag spores to agar? [Re: Psilicon] * 2
    #26843893 - 07/25/20 01:23 AM (3 years, 6 months ago)

Yeah, later zones have  less spores, less contaminates, less strains present in each colony


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C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche


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Offlinejcm4620
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Re: Why zig-zag spores to agar? [Re: c10h12n2o] * 1
    #26843963 - 07/25/20 03:47 AM (3 years, 6 months ago)

i just figured this would be a good place to show off this great red spored pan aussie i got. fucking 36hours in i made 10 transfers per plate. these pics were takin at the 48 hour mark and you can hardly tell in some spots. i have never had this kind of growth so fast  from a pan print.



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PANAEOLUS FRUITING MADE SIMPLE



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OfflinePsilicon
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Re: Why zig-zag spores to agar? [Re: jcm4620]
    #26844033 - 07/25/20 05:58 AM (3 years, 6 months ago)

That’s gorgeous. You do three times around with the parafilm?

Do it like TC Moneybags over here if you can, OP :rofl:


--------------------
Agar - what, why and how?  Everything a beginner needs to know.
Oat Prep Tek
Bored?  Please take one of my experiments off my hands.


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Offlinejcm4620
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Re: Why zig-zag spores to agar? [Re: Psilicon]
    #26844049 - 07/25/20 06:13 AM (3 years, 6 months ago)

lol not sure how many times i go around lol i just use 3 squares so whatever that is. a fucking roll of para lasts me a long ass time.


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PANAEOLUS FRUITING MADE SIMPLE



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OfflineLogicaL ChaosM
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Re: Why zig-zag spores to agar? [Re: maxmush]
    #26844513 - 07/25/20 12:00 PM (3 years, 6 months ago)

Quote:

maxmush said:
I understand this is common practice and I have done myself, but what exactly is the rationale?




It helps isolate strains and infections (containments) which makes them easier to work with, as already mentioned above :thumbup:


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OfflineC12ShroomMan
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Re: Why zig-zag spores to agar? [Re: LogicaL Chaos] * 1
    #26844522 - 07/25/20 12:04 PM (3 years, 6 months ago)

Z for ZORO, protector of mycelium





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OfflineLogicaL ChaosM
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Re: Why zig-zag spores to agar? [Re: C12ShroomMan]
    #26844524 - 07/25/20 12:05 PM (3 years, 6 months ago)

Your Z's are backwards :datass:


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OfflineC12ShroomMan
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Re: Why zig-zag spores to agar? [Re: LogicaL Chaos]
    #26844530 - 07/25/20 12:06 PM (3 years, 6 months ago)

Flip upside down or look in mirror ' S ' is only an illusion :crazy2:


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Invisiblemaxmush
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Registered: 06/13/20
Posts: 440
Re: Why zig-zag spores to agar? [Re: C12ShroomMan]
    #26845012 - 07/25/20 06:10 PM (3 years, 6 months ago)

Does anyone have some pics of the optimal sectoring areas on a "streak"? Many thanks!


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Disclaimer: all information presented is intended for educational purposes only. All photos are only representations and not directly from the user.


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Offlinejcm4620
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Re: Why zig-zag spores to agar? [Re: maxmush] * 1
    #26845108 - 07/25/20 07:50 PM (3 years, 6 months ago)

i think zoro may have been asleep cuz im not sure if thats even myc. or atleast what i call myc if its cubes ur lookin for heres a few examples




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OfflineCamera93
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Re: Why zig-zag spores to agar? [Re: jcm4620]
    #26845283 - 07/25/20 10:28 PM (3 years, 6 months ago)



Hes asking about from a streak plate
This is cube myc here germinated from spore


--------------------
All I need are some tasty waves, a cool buzz, and I’m fine.

Whatever you decide won’t really impact our survival
Close your eyes, and do the best that you can



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OfflineC12ShroomMan
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Re: Why zig-zag spores to agar? [Re: jcm4620]
    #26845285 - 07/25/20 10:29 PM (3 years, 6 months ago)

zoro be swashblucklin. heres transfers from that plate if adventure be what ye be searchn for





Edited by C12ShroomMan (07/25/20 10:31 PM)


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Offlinejcm4620
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Re: Why zig-zag spores to agar? [Re: C12ShroomMan]
    #26845472 - 07/26/20 03:32 AM (3 years, 6 months ago)

def looking better


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