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OfflineExcel89
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Registered: 08/27/13
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First time innoculating agar/petri dishes, is this normal?
    #18791777 - 09/02/13 08:06 PM (10 years, 5 months ago)

I used 10ccs of a spore syringe that I made to inoculate about 30 petri dishes. I'm not sure if that's too little per dish, but this is my first time dealing with agar so I'm just testing the waters and didn't want to waste too much in case I did something wrong. This is what they're starting to look like after 2 days. Is this normal? The agar is turning darker in the areas where I inoculated them, but I don't see any mycelium yet. How long should I expect it to take? Or did I do something wrong and is this just mold/contamination/water? The petri dishes I bought from amazon don't completely seal either, so I'm aware that I may have to start over and use different petri dishes, I just want to make sure I know what I'm doing before I go that route.

Any advice would be appreciated.



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OfflineKizzle
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Excel89]
    #18791979 - 09/02/13 08:52 PM (10 years, 5 months ago)

There's too much water there. I would try to avoid inoculating agar with a syringe altogether although it is possible. What you really want is a spore print so you can transfer the dry spores to the agar.


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OfflineExcel89
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Kizzle]
    #18791996 - 09/02/13 08:58 PM (10 years, 5 months ago)

Well I tried both. I have another 10 with spore prints, but I haven't seen much growth and it's been a week and a half. I'll take a picture in a bit, but I see the physical spores on the agar, and a couple of them have a little bit of what looks like mycelium, but it's very weak. Someone told me that using a syringe on agar causes faster growth which is why I tried it that way. =/


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Invisiblemushmagic
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Kizzle]
    #18792003 - 09/02/13 08:59 PM (10 years, 5 months ago)

:whathesaid: and if you do inoculate agar with a syringe you only want to use a drop or two max. As kizzle said though just go straight from the print next time


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Offlinevec

Registered: 02/26/13
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Excel89]
    #18792026 - 09/02/13 09:05 PM (10 years, 5 months ago)

Those dishes in the picture are all contaminated! Throw them out, start again. Make sure you have a Glovebox and a print or live tissue. Sterility is high priority when it comes to dishes. You still have a long way to go young Jedi.


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OfflineExcel89
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Excel89]
    #18792142 - 09/02/13 09:26 PM (10 years, 5 months ago)

Thanks guys. I posted this in another thread but I actually used prints that I took about 6 months-10 months ago. I took some prints, but them between two pieces of clear laminate paper, put jars over them, and forgot about them. I randomly came across them and wanted to see if I could save them, so I made 10 spore syringes, put the rest of the prints into petri dishes, and save one of the prints. I wanted to see if I could get anything clean out of it, which is why I used the agar in the first place knowing they were contaminated.

I guess I'll have to just buy some syringes online and start from scratch. :P

Do you guys think the petri dishes I'm using are ok? I have another 70 of them, but they aren't the greatest. The lids just rest on top they don't fully close. Or do I need to buy new petri dishes, new syringes/prints, and start from scratch working in a glove box?


Edited by Excel89 (09/02/13 09:27 PM)


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Offlinevec

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Re: First time innoculating agar/petri dishes, is this normal? [Re: Excel89]
    #18792167 - 09/02/13 09:31 PM (10 years, 5 months ago)

Looks like you are getting a little ahead of yourself. Start with syringes and grains. Then do some grain to grain transfers. Then actually grow some and use some live tissue or spore print from fresh mush to clone some.


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Offlinevec

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Re: First time innoculating agar/petri dishes, is this normal? [Re: vec]
    #18792192 - 09/02/13 09:34 PM (10 years, 5 months ago)

those dishes look plastic with vented tops. vented tops are good, plastic dishes come pre-sterilized in the bag. If you dont use all of them at once I would suggest throwing them out.


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OfflineExcel89
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Excel89]
    #18792233 - 09/02/13 09:40 PM (10 years, 5 months ago)

I actually have quite a lot of experience with growing. Mainly I would use BRF substrate in pint jars, make my own syringes by washing the sides of the jars with a syringe and sucking it back up, and fruit them in a storage tote. I'd put clear plexiglass on top, made a drip shield using saran wrap, etc. The problem is I've always made my own from syringes from already colonized brf cakes, I have absolutely no experience with spore prints or agar, but I decided to give it a shot because I'm trying to find a way to get into a full scale thing without waiting two months to colonize the jars the way I normally do. I'll probably order some syringes online since mine must be contaminated, and I'll inoculate some jars and use a little to play with petri dishes some more. I know what I'm doing when it comes to syringes and BRF, just not with prints and agar. Thanks for the help guys.


Edited by Excel89 (09/02/13 09:40 PM)


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OfflineMr.man
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Excel89]
    #24240020 - 04/13/17 10:35 AM (6 years, 10 months ago)

Quote:

Excel89 said:
I used 10ccs of a spore syringe that I made to inoculate about 30 petri dishes. I'm not sure if that's too little per dish, but this is my first time dealing with agar so I'm just testing the waters and didn't want to waste too much in case I did something wrong. This is what they're starting to look like after 2 days. Is this normal? The agar is turning darker in the areas where I inoculated them, but I don't see any mycelium yet. How long should I expect it to take? Or did I do something wrong and is this just mold/contamination/water? The petri dishes I bought from amazon don't completely seal either, so I'm aware that I may have to start over and use different petri dishes, I just want to make sure I know what I'm doing before I go that route.

Any advice would be appreciated.






To much nute, I believe. You have to dilute your soup with more water,  you can tell because you can't see through your agar. Look at pictures of how your agar should look. I'm new but I believe that sappy ooze is from to much nute, plus if u want fast colonization you make your agar light not thick.  Correct me if I'm wrong,  I'm still a student as well


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InvisiblebodhisattaMDiscordReddit
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Mr.man]
    #24240029 - 04/13/17 10:36 AM (6 years, 10 months ago)

10cc syringe can inoculate a few hundred dishes probably a thousand or more

Anything more than a single drop is too much. I use Less than one drop per plate


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OfflineMr.man
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Excel89]
    #24240030 - 04/13/17 10:36 AM (6 years, 10 months ago)

Quote:

Excel89 said:
I used 10ccs of a spore syringe that I made to inoculate about 30 petri dishes. I'm not sure if that's too little per dish, but this is my first time dealing with agar so I'm just testing the waters and didn't want to waste too much in case I did something wrong. This is what they're starting to look like after 2 days. Is this normal? The agar is turning darker in the areas where I inoculated them, but I don't see any mycelium yet. How long should I expect it to take? Or did I do something wrong and is this just mold/contamination/water? The petri dishes I bought from amazon don't completely seal either, so I'm aware that I may have to start over and use different petri dishes, I just want to make sure I know what I'm doing before I go that route.

Any advice would be appreciated.






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OfflineMr.man
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Excel89]
    #24240034 - 04/13/17 10:37 AM (6 years, 10 months ago)

Quote:

Excel89 said:
I used 10ccs of a spore syringe that I made to inoculate about 30 petri dishes. I'm not sure if that's too little per dish, but this is my first time dealing with agar so I'm just testing the waters and didn't want to waste too much in case I did something wrong. This is what they're starting to look like after 2 days. Is this normal? The agar is turning darker in the areas where I inoculated them, but I don't see any mycelium yet. How long should I expect it to take? Or did I do something wrong and is this just mold/contamination/water? The petri dishes I bought from amazon don't completely seal either, so I'm aware that I may have to start over and use different petri dishes, I just want to make sure I know what I'm doing before I go that route.

Any advice would be appreciated.






To much nute, I believe. You have to dilute your soup with more water,  you can tell because you can't see through your agar. Look at pictures of how your agar should look. I'm new but I believe that sappy ooze is from to much nute, plus if u want fast colonization you make your agar light not thick.  Correct me if I'm wrong,  I'm still a student as well


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InvisiblebodhisattaMDiscordReddit
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Mr.man]
    #24240036 - 04/13/17 10:38 AM (6 years, 10 months ago)

Yea don't triple post the same thing.


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OfflineCapnZ
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Re: First time innoculating agar/petri dishes, is this normal? [Re: bodhisatta]
    #24240046 - 04/13/17 10:43 AM (6 years, 10 months ago)

Quote:

bodhisatta said:
10cc syringe can inoculate a few hundred dishes probably a thousand or more

Anything more than a single drop is too much. I use Less than one drop per plate



:whathesaid:

I'm still getting the " less than a drop" technique down, but I've been getting decent results with just one drop of MS per plate.


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InvisiblebodhisattaMDiscordReddit
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Re: First time innoculating agar/petri dishes, is this normal? [Re: CapnZ]
    #24240063 - 04/13/17 10:50 AM (6 years, 10 months ago)

I use an inoculation loop


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OfflineShroominmojo0213
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Excel89]
    #24240253 - 04/13/17 11:54 AM (6 years, 10 months ago)

Quote:

Excel89 said:
Thanks guys. I posted this in another thread but I actually used prints that I took about 6 months-10 months ago. I took some prints, but them between two pieces of clear laminate paper, put jars over them, and forgot about them. I randomly came across them and wanted to see if I could save them, so I made 10 spore syringes, put the rest of the prints into petri dishes, and save one of the prints. I wanted to see if I could get anything clean out of it, which is why I used the agar in the first place knowing they were contaminated.

I guess I'll have to just buy some syringes online and start from scratch. :P

Do you guys think the petri dishes I'm using are ok? I have another 70 of them, but they aren't the greatest. The lids just rest on top they don't fully close. Or do I need to buy new petri dishes, new syringes/prints, and start from scratch working in a glove box?




If the prints were not dry, as in you left the caps on them too long than they will be bacterial


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InvisiblebodhisattaMDiscordReddit
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Shroominmojo0213]
    #24240258 - 04/13/17 11:55 AM (6 years, 10 months ago)

You can leave the caps on any amount of time. You always have to let a print dry before folding it up anyway


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InvisibleMoabfighter
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Re: First time innoculating agar/petri dishes, is this normal? [Re: bodhisatta]
    #24240260 - 04/13/17 11:56 AM (6 years, 10 months ago)

Quote:

bodhisatta said:
I use an inoculation loop




Squirt spore drop on loop, rub loop on agar?

I've tried spore water to agar like 5-8 times. Never gotten germination. Tried the middle, tried rolling the drop around the outer circle of the plate, nothing.

I do have a loop, and think that may be a great idea. Maybe stab the agar with the loop so it gets nice and deep inside?


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InvisiblebodhisattaMDiscordReddit
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Re: First time innoculating agar/petri dishes, is this normal? [Re: Moabfighter]
    #24240265 - 04/13/17 11:57 AM (6 years, 10 months ago)

The methodology is in my agar tek with a gif image too


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