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OfflineBackbone
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Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water?
    #26486966 - 02/15/20 11:49 AM (3 years, 11 months ago)

sorry its worded so weird


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OnlineLogicaL ChaosM
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Backbone]
    #26486994 - 02/15/20 12:10 PM (3 years, 11 months ago)

Got a photo?

U shouldnt have streak marks from shooting spore solution on agar.


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OfflineMr.Wizard
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: LogicaL Chaos]
    #26486997 - 02/15/20 12:12 PM (3 years, 11 months ago)

My first plates I got too much spore solution on them, and as the water dried I could see where it had traveled around the plate. On other plates that I used in inoculation loop on in the right light I could see my streak marks.

Pics would be very helpful.


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OfflineBackbone
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Mr.Wizard]
    #26487004 - 02/15/20 12:16 PM (3 years, 11 months ago)

id take a picture but itd be hard to see as its in my sab


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OfflineMr.Wizard
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Backbone] * 1
    #26487011 - 02/15/20 12:20 PM (3 years, 11 months ago)

It sounds like you should do some more research, I wouldn't worry about the streaks. Read through the middle link in my sig at least. The bottom link has some good info for agar too.

Once you have your plate made it's able to come out of the SAB. I'm not sure why that's not allowing to take a picture. If it's just sitting open in there it's probably trash now. Here's where I put mine:



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OfflineBackbone
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Mr.Wizard]
    #26487016 - 02/15/20 12:23 PM (3 years, 11 months ago)

What why would it be trash


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OfflineMr.Wizard
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Backbone]
    #26487043 - 02/15/20 12:46 PM (3 years, 11 months ago)

There are still contams in an SAB, and having the container open means they can get in. Things should be exposed to open air/be open, as little time as possible, even in the SAB.  That's good sterile technique, and what I would recommend researching specifically.


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OfflineBackbone
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Mr.Wizard]
    #26487072 - 02/15/20 01:13 PM (3 years, 11 months ago)

i didnt feel like fing with wrapping each dish even though i only made three so i just wrapped an arm hole i guess if there's contams the mycelium wont grow into it? but other than what im hoping is just dissolved syringe water on the surface, they look pretty good. id take a picture but i cant find my phone i don't even think you'd see it unless i exposed the dishes to open air


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OfflineMr.Wizard
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Backbone]
    #26487099 - 02/15/20 01:33 PM (3 years, 11 months ago)

Well you do you, but the folks here were gracious enough to give us this info that has been proven to work. Anything I do because I feel like it is in addition to, and after doing everything by their book.

Like I said, the possible streaking from the syringe liquid is probably not your biggest concern.

Without having any information about dates of inoculation, or pics of your stuff there's not a lot folks can tell you.

I will say that if they have just been in open air in your SAB since inoculation they probably aren't growing because they are in extremely oxygenated air, and probably entirely contaminated.

Oxygen (FAE) is for fruiting. A CO2 rich environment is what they need right now, as if they were underground. There is tons of fresh air in your SAB.

Honestly I say you need to put in your due diligence of several hours of research over some days, if not weeks. Then restart while following known good techniques.

Sorry if that's a little harsh, but it's very nearly a waste of all our times to entertain this discussion further if your plates have just been sitting open in your SAB for any extended amount of time. Very nearly.


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Offlinemimir
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Backbone]
    #26487114 - 02/15/20 01:44 PM (3 years, 11 months ago)

If you can see the water from the syringe standing still in one place, it's most likely bacterial contamination. If not, water will either evaporate or keep sloshing around, carrying around any contaminants that might be usually at the edge of the dish. That is why it's generally preferred to start from a print if you're doing agar, or else use as little solution from the syringe as possible.


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Invisiblealaskappalachian
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: mimir]
    #26487135 - 02/15/20 01:57 PM (3 years, 11 months ago)

Yeah if you put too much ms solution on your plate you generally get bacterial snot streaks and weird growth. One drop in a couple/few places is great.  Producing A drop is difficult, but not impossible.  You can pour a thick plate and stab into it, then as you are withdrawing, sort of try to depress a drop into the recess created.  It's worth wasting a few drops (if you're having trouble) to get a drop formed on the end of your needle directly beside your open plate, then carefully bring the needle over to the agar to avoid an acidental squirt.


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Offlinemimir
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: alaskappalachian]
    #26487141 - 02/15/20 02:06 PM (3 years, 11 months ago)

Another way is to use an inoculation loop. Squirt some MS solution into a clean plate with no agar then dip your inoculation loop and streak


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OfflineBackbone
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: alaskappalachian]
    #26487153 - 02/15/20 02:13 PM (3 years, 11 months ago)

i meannnnn it looks fine, plenty of inoculation points, wizard i dont think you know what youre talking about because there is bacteria that lives off of no oxygen and mycelium in the wild doesn't grow only underground the top layer is constantly receiving fresh air


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OfflineBackbone
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: mimir]
    #26487162 - 02/15/20 02:18 PM (3 years, 11 months ago)

Yeah i was an idiot and i didnt fully understand what to do with the inoculation loop so i squirted on to the plate first then was like uhhhh i guess i make a zigzag. It just seems weird to me that i could get enough spores on my loop to do enough transfers before the mycelium died out but i guess i should trust these guys' experience


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Offlinemimir
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Backbone]
    #26487181 - 02/15/20 02:32 PM (3 years, 11 months ago)

If I'm being honest, last time I did agar I just squirted into the Petris with little to no care. Yes, there was water swimming around, yes I think I got like one or two bacterial contamination. If the syringe is good, outside contaminants, even with plenty of water swimming around, are mostly a matter of skill. Even if you do get some outside contamination, as long as you have one part that shows growth you can just transfer, that's the whole point of agar. If you only had 2 plates to inoculate, then yes: inoculation loop or a single drop, flow hood and so on. But honestly, just inoculate like 6 plates or more, all you need is one good speck :shrug:
If that then fails, and the syringe is so dirty that you get contamination everywhere no matter how good the technique, just get a refund, one drop or ten drops won't make the difference there.


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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: mimir]
    #26487194 - 02/15/20 02:41 PM (3 years, 11 months ago)

I honestly don't think anyone on this thread has a clue what their are talking about.

Wizard, can you explain what you mean when you say "a CO2 rich environment is what "they" need right now, as if they were underground?"

Backbone, I like to use my inoculation loop to spread spores when I transfer off of a print. When using syringe, just squeeze out a little drop and streak it with the tip of the needle like you would an inoculation loop. Also you should probably go a head and wrap the dish edge with Saran Wrap, but it is up to you. If you inoculated a dish and it shows a raised jelly area where your water was its probably bacterial contam, but it is hard to say with out pictures.


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OfflineMr.Wizard
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Sockadin]
    #26487205 - 02/15/20 02:50 PM (3 years, 11 months ago)

Meaning that they need an environment that is not fresh air, with only some gas exchange.

While we grow myceleum we are trying to recreate their natural environment, which is to grow underground until myceleum hits fresh air at the top of the dirt, then fruit.

His plates might as well be in open air if my understanding that they are in a sealed SAB without lids on is correct. I think talking about anything else is a band-aid on a bullet wound at this point.


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I am so happy and grateful that we get to live in joyful abundance, while things get better, and better.


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Offlinemimir
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Mr.Wizard]
    #26487228 - 02/15/20 03:06 PM (3 years, 11 months ago)

Quote:

Mr.Wizard said:
Meaning that they need an environment that is not fresh air, with only some gas exchange.

While we grow myceleum we are trying to recreate their natural environment, which is to grow underground until myceleum hits fresh air at the top of the dirt, then fruit.

His plates might as well be in open air if my understanding that they are in a sealed SAB without lids on is correct. I think talking about anything else is a band-aid on a bullet wound at this point.



He doesn't have the dishes open, they are just not wrapped. They will get GE, wrapped or not, considering that in labs they are usually wrapped with parafilm, which allows for GE. Having them not wrapped shouldn't be much of a problem if they're just sitting still inside of the SAB. That's how Petris are often kept in incubators in microbiology labs, sometimes they're not even wrapped. Sure, if you want to be able to move them around easily and just have peace of mind, you should by all means wrap them, but otherwise the plates are designed to make it hard for stuff to get in once they're closed.


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OfflineBackbone
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: Mr.Wizard]
    #26487234 - 02/15/20 03:14 PM (3 years, 11 months ago)

what lol theres no air currents in my sab, aside from possible temperature differences true it'd be better to wrap them, probably. But i just want to point out to you that what youre saying is wrong, are you familiar with pf tek? its off topic but im just saying, this is to wizard. Also im thinking fuck streaking and just put one drop in the center of the agar dish


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OfflineMr.Wizard
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Re: Does anyone know, i got streak marks on my agar plates i used a ms syringe, is it from the water? [Re: mimir]
    #26487239 - 02/15/20 03:18 PM (3 years, 11 months ago)

Ohh! Ok, I was horrified at the thought of them just being open in there, lol.

But I stand by what I said about the streaking, don't worry about it. I saw streaking on both the below dishes (MS syringe inoculation the same day) I only had growth on one, but the other you can still see the streak from my inoculation loop. It's in the middle of the dish, you can see the line through the light.



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I am so happy and grateful that we get to live in joyful abundance, while things get better, and better.


Edited by Mr.Wizard (02/15/20 03:22 PM)


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