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OfflineNightPuma1
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Agar from MS syringe question
    #26637162 - 04/29/20 04:24 PM (3 years, 8 months ago)

I would like sort of a vote if possible on the preferred method BETWEEN THESE TWO (I know a lot of people do streaking). Using a MS syringe on agar:

1. 3 drops on the petri dish in a triangular pattern

2. 1 drop in the center, followed by tilting the dish slightly causing the drop to run towards the edge (but not touching it).

Thank you!


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OfflineHouseOfRoses
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Re: Agar from MS syringe question [Re: NightPuma1] * 1
    #26637168 - 04/29/20 04:25 PM (3 years, 8 months ago)

I havent actually used agar with an MS syringe yet, but I was always told just one drip in the middle and no tilting the dish


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Re: Agar from MS syringe question [Re: HouseOfRoses] * 1
    #26637171 - 04/29/20 04:28 PM (3 years, 8 months ago)

#2 if I had to choose between the 2 :shrug:


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OfflineNightPuma1
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Re: Agar from MS syringe question [Re: Tormato]
    #26637373 - 04/29/20 06:16 PM (3 years, 8 months ago)

If I may ask, what is the actual advantage in introducing a new potential contamination vector (meaning the inoculation loop) just to get the droplet to do a zig-zag pattern?


Edited by NightPuma1 (04/29/20 06:17 PM)


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Re: Agar from MS syringe question [Re: NightPuma1] * 3
    #26637378 - 04/29/20 06:19 PM (3 years, 8 months ago)

To spread it out which won't be a vector if you flame sterilize the loop first.

Honestly the MS syringe is gonna be your biggest potential vector


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Re: Agar from MS syringe question [Re: Tormato] * 2
    #26637466 - 04/29/20 07:02 PM (3 years, 8 months ago)

:whathesaid:

When you streak the spores you should see germination in the zig zag pattern of the streak and have a much easier time distinguishing clean germination from things like mold colonies + bacteria.

If you are going to be working with agar, you are going to have to get used to opening the lid and using tools like inoculation loops and scalpels to swipe and make transfers. 


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Edited by mushpunx (04/30/20 02:42 AM)


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OfflineLogicaL ChaosM
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Re: Agar from MS syringe question [Re: NightPuma1]
    #26637761 - 04/29/20 09:56 PM (3 years, 8 months ago)

I vote one drip in the center, no tilting. Dont forgot to shake the syringe first !


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Re: Agar from MS syringe question [Re: LogicaL Chaos] * 1
    #26637828 - 04/29/20 10:39 PM (3 years, 8 months ago)

Quote:

LogicaL Chaos said:
I vote one drip in the center, no tilting. Dont forgot to shake the syringe first !



:thumbup:


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Re: Agar from MS syringe question [Re: tripdawg420]
    #26637835 - 04/29/20 10:43 PM (3 years, 8 months ago)

:awebig:


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Re: Agar from MS syringe question [Re: LogicaL Chaos] * 1
    #26637847 - 04/29/20 10:53 PM (3 years, 8 months ago)

Assuming it's a well shaken syringe, as little as possible, one drop in the center with as little liquid as possible seems the best to me. just my 2¢


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Offlinepoisoned
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Re: Agar from MS syringe question [Re: mistermushly]
    #26638050 - 04/30/20 01:14 AM (3 years, 8 months ago)

I used to do a drop in the center. Streaking is 10x better.


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Re: Agar from MS syringe question [Re: poisoned] * 2
    #26638298 - 04/30/20 05:25 AM (3 years, 8 months ago)

The draw back to leaving the drop in the center is that now everything is going to grow in a big mass ..contams included.

Always best to spread it out for visibility reasons.


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OfflineLogicaL ChaosM
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Re: Agar from MS syringe question [Re: Tormato]
    #26638369 - 04/30/20 06:44 AM (3 years, 8 months ago)

Can u streak with a syringe needle or do u need a Inocu loop?


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Re: Agar from MS syringe question [Re: LogicaL Chaos] * 1
    #26638444 - 04/30/20 07:33 AM (3 years, 8 months ago)

Quote:

LogicaL Chaos said:
Can u streak with a syringe needle or do u need a Inocu loop?



You can, but I find a loop just spreads it better


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Offlinepoisoned
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Re: Agar from MS syringe question [Re: Tormato] * 1
    #26638453 - 04/30/20 07:38 AM (3 years, 8 months ago)

I tried to, but it's a bit more tricky. You can make a loop from materials you probably have at home in a minute.


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Offlinetripdawg420
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Re: Agar from MS syringe question [Re: Tormato] * 1
    #26638552 - 04/30/20 08:40 AM (3 years, 8 months ago)

Quote:

Tormato said:
The draw back to leaving the drop in the center is that now everything is going to grow in a big mass ..contams included.

Always best to spread it out for visibility reasons.



contams and myc wont mix together:shrug:


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OfflineThe Mycologist
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Re: Agar from MS syringe question [Re: tripdawg420]
    #26638569 - 04/30/20 08:53 AM (3 years, 8 months ago)

Quote:

tripdawg420 said:
Quote:

Tormato said:
The draw back to leaving the drop in the center is that now everything is going to grow in a big mass ..contams included.

Always best to spread it out for visibility reasons.



contams and myc wont mix together:shrug:




Yes they can


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Re: Agar from MS syringe question [Re: The Mycologist]
    #26638573 - 04/30/20 08:55 AM (3 years, 8 months ago)

Quote:

The Mycologist said:
Quote:

tripdawg420 said:
Quote:

Tormato said:
The draw back to leaving the drop in the center is that now everything is going to grow in a big mass ..contams included.

Always best to spread it out for visibility reasons.



contams and myc wont mix together:shrug:




Yes they can



oh ya plz explain


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coir tek
http://www.shroomery.org/forums/showflat.php/Number/11917410
results :thumbup:


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Re: Agar from MS syringe question [Re: tripdawg420] * 1
    #26638660 - 04/30/20 10:01 AM (3 years, 8 months ago)

You definitely want to take transfers of myc that haven't grown into/over or collided with bacteria/mold colonies.

I have had bacterial cultures before , they get this strange puffy look (not tomentose). Man it's hard to explain I wish I still had some photos


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Re: Agar from MS syringe question [Re: mushpunx] * 1
    #26638674 - 04/30/20 10:08 AM (3 years, 8 months ago)

:whathesaid: Right...it's just easier to discern what's what and what hasn't been possibly compromised when you spread things out :shrug:


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OfflineThe Mycologist
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Re: Agar from MS syringe question [Re: tripdawg420] * 1
    #26638675 - 04/30/20 10:09 AM (3 years, 8 months ago)

I have had both molds and bacteria riding along with a culture.

They dont always act like there is a barrier.

Streaking comes from actual laboratory practice, you think they do it for no reason?


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OfflineNightPuma1
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Re: Agar from MS syringe question [Re: The Mycologist]
    #26640028 - 04/30/20 09:54 PM (3 years, 8 months ago)

No, it just seems to me that if you have 2 methods of spreading out a drop and one of them does not involve sticking something in there then that was the way to go. Apparently I am mistaken - I just haven't really gotten a great answer of why exactly that is.  Why a zig-zag pattern is better than the streak I get by letting it slide. I believe you I just don't understand.


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Re: Agar from MS syringe question [Re: NightPuma1]
    #26640052 - 04/30/20 10:11 PM (3 years, 8 months ago)

How about a 3rd option, put a couple drops on a sterile swab and streak with that.


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Re: Agar from MS syringe question *DELETED* [Re: SYF8] * 1
    #26640197 - 05/01/20 12:13 AM (3 years, 8 months ago)

Post deleted by Grenade01

Reason for deletion: .


Edited by Grenade01 (05/01/20 12:14 AM)


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OfflineLogicaL ChaosM
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Re: Agar from MS syringe question [Re: Grenade01]
    #26640205 - 05/01/20 12:19 AM (3 years, 8 months ago)

I hate when i shoot too much spores :awehigh:


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Re: Agar from MS syringe question [Re: LogicaL Chaos]
    #26640252 - 05/01/20 12:44 AM (3 years, 8 months ago)

How would introducing too many spores causes issues?


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Re: Agar from MS syringe question [Re: MrMong]
    #26640261 - 05/01/20 12:50 AM (3 years, 8 months ago)

It makes it harder to seperate good mycelium from contaminates. Since its all microscopic, it will make it a pain in the ass to cut sections off. Thats why streaking is preferred because it spreads the spores over a wider area in the agar, thus making sectoring easier.


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Re: Agar from MS syringe question [Re: NightPuma1]
    #26640296 - 05/01/20 01:17 AM (3 years, 8 months ago)

Quote:

NightPuma1 said:
No, it just seems to me that if you have 2 methods of spreading out a drop and one of them does not involve sticking something in there then that was the way to go. Apparently I am mistaken - I just haven't really gotten a great answer of why exactly that is.  Why a zig-zag pattern is better than the streak I get by letting it slide. I believe you I just don't understand.




You flame sterilize that thing, it shouldn't be an issue at all.


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Offlinemushpunx
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Re: Agar from MS syringe question [Re: NightPuma1] * 1
    #26640524 - 05/01/20 05:27 AM (3 years, 8 months ago)

Prints are never 100% clean, and therefore spore syringes are not either.
Unless you have sloppy technique, contamination on the plate is a lot more likely to grow from the spore solution that be introduced by a sterilized loop.

When you let that spore solution slide around the plate, any potential mold colonies (which are often white and resemble mushroom myc) and bacteria (which can be blobs- or a light film that is very hard to spot) will grow in the same area as the germination, they might touch or the myc might grow over or into it. It doesn't mean you won't be able to transfer some clean myc away from it, but when you swipe the spores in a zig zag pattern across the plate, first you know that the germination should occur in that pattern and not to transfer from anything outside of the zig zag, and second it spreads the spores out so that you might more easily be able to take a transfer of clean myc that is far away from mold or bacteria.

You don't have to do this, it's just good technique. If you are going to be working with agar you are going to have to learn to use sterile tools


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Edited by mushpunx (05/01/20 05:28 AM)


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OfflineThe Mycologist
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Re: Agar from MS syringe question [Re: NightPuma1]
    #26640605 - 05/01/20 06:40 AM (3 years, 8 months ago)

Quote:

NightPuma1 said:
No, it just seems to me that if you have 2 methods of spreading out a drop and one of them does not involve sticking something in there then that was the way to go. Apparently I am mistaken - I just haven't really gotten a great answer of why exactly that is.  Why a zig-zag pattern is better than the streak I get by letting it slide. I believe you I just don't understand.




You have to have faith in the techniques.

This whole thing relies on flame sterilization working how its intended.

But If you want to limit your vectors, that has logic too, and may be good for someone starting out.

...

I was originally responding to the guy who was saying that contams cant grow along with mycelium.


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That the powerful play goes on, and you will contribute a verse.”
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