first crack at petri dishes ( with pics)

Discussion in 'MUSHROOM CULTIVATION' started by nomendubium, Feb 29, 2012.

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  1. nomendubium

    nomendubium scraping by, since '97 Expert Identifier

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    so, I poured my disposable petri dishes yesterday.
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    they are the 70mm variety. I traded some of my pics for them:dance:
    So, I added 18grams +- malt extract agar to 360ml of cool tap water, brought it to a simmer for 10 minutes, put it in a pint jar and pc'ed 30 minutes @10psi.
    when it was cool, I sprayed everything with oust and loaded the sab.
    I did the old'"lift the stack" method to pour them, and then I left them in stacks of 5 to cool. After a couple hours of cooling, they were solid and I wrapped them with parafilm. I know I know, I wasted the parafilm, but heres how I look at it; I had enough to wrap them 2 times and have never used it before. You don't take a new gun in the woods to hunt, if you've never shot it before, so you're gonna have to fire a few rounds to get a feel for i:dance:. I'm glad I did :wink1:
    Out of the 20 dishes, I have 14 that I feel good about. I have 3 that the lid seperated from the base while trying to wrap them, and 3 more I busted with finger-pressure while trying to wrap.
    If I hadn't tried it out, I would have probably busted them after I transfered a culture, and was trying to wrap it, so I probably saved myself a head-ache there.
    It was actuallt pretty damn difficult, starting off, but by the time I got to the end, It was rolling along smoothly, so now I have a little experience and a little more confidence to move ahead. I'll give them a few days and see what they do, if they are contaminated or not, and I'll start using them. Oh, yea, I said pics.
    [​IMG]
    here they are, on my messy-ass dresser
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    one from the top
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    side veiw. I didn't pick the best one to photo, I just picked one off the top, so some look better/more evenly wrapped
    my verdict
    it was pretty damn hard, but hopefully I get better at it with practice :press:
     
  2. CharlieBrown

    CharlieBrown Member

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    You will DEFINITELY get better at it with practice. It felt awkward for me the very first go at it, but you get used to what you are working with.

    You'll be surprised, I too have set aside ones that the bottoms came apart from the top for whatever reason after pouring and have had no issues with contams. I always thought if you even look at agar dishes wrong the media will contam, however turns out if you are good on your sterile practices you will probably be ok.
     
  3. nomendubium

    nomendubium scraping by, since '97 Expert Identifier

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  4. Vibration

    Vibration Active Member

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    Wow, is it really important to wrap them while they're cooling? What does it help? Does it help with the mold spores or bacteria?

    I have never had any mold spores getting into my agar plates while they're cooling.

    I really need to know if it's for mold spores or also for bacteria.

    Hahaha.
     
  5. nomendubium

    nomendubium scraping by, since '97 Expert Identifier

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    no? I didn't wrap them while they were cooling, I just wrapped them after they were cool, but before I took them out of the sab. Most people suggest wrapping them in saran wrap, But I only had an old dusty roll and I didn't feel comfortable using dirty saran wrap to wrap them in
    I have a bad habit of not cleaning, and it wouldn't be a problem if I could just leave them setting still, but I have limited space and also wanted to gain some parafilm experience. At the rate I do mycology, I may never have agar petri dishes and parafilm at the same time again, so I wanted to make my first attempt a really good one. If you don't have problems with your dishes, there is no good reason to wrap them.
     
  6. Vibration

    Vibration Active Member

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    Thank you for the response and sorry for my little mis-understanding :)
     
  7. nomendubium

    nomendubium scraping by, since '97 Expert Identifier

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    no problemo! Perhaps stay tuned, if you are lucky/I am unlucky you can watch me fail miserably and that will boost everyones mood. Lately I've done a lot of talking and not a lot of doing, so I'm trying to change that now. Also these are my first attempts at the new place(which is probably cleaner than the last few places I've been)
     
  8. Vibration

    Vibration Active Member

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    An unluck of yours won't make me lucky :D believe me, I don't want to see you failing. Considering how many helpful postings you've made on my thread, I think you are going to be just fine. I think you know what you're doing.
     
  9. nomendubium

    nomendubium scraping by, since '97 Expert Identifier

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    we'll find out, before long. If I can get agar down, I feel good about my grain prep, bulk subs ect. It's really been a stumbling block, not having experience with it because without it, a spore print is pretty much either good or bad, and if it's bad there is no saving it, but using agar opens a lot of doors in that hallway
     
  10. Vibration

    Vibration Active Member

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    Yes, I want to believe this is how it is.
     
  11. nomendubium

    nomendubium scraping by, since '97 Expert Identifier

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    bad news, guys. apparently something went wrong. Look at these pics, are they absolutely swimming in contamination or is there another explaination?
    ZDo you think I didn't pc the agar long enough? (1/2 hour @10psi) or do you think the dishes were contaminated, or do you think my sab was absolutely full of contamination? I sprayed neutra-air, lysoled everything that went in and didn't have them open for any excess time at all
     

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  12. CharlieBrown

    CharlieBrown Member

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    Ouch. I do mine for 40 minutes at 15psi. 10psi for 30 mins seems low
     
  13. rogue

    rogue ♥ Hooked on Mycelium ♥ Moderator Mushroom Doctor Supporter

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    How did you pc the agar? Did you have it in a container with a filter. Looking at the contam pattern It looks like you need to PC longer.

    You might be able to get away with it doing PP no-pour method as the jars have little agar in them but doing 500 - 1000 mls would require more time.
     
  14. Professor PinHead

    Professor PinHead Lost in the Tek.... Administrator Mushroom Doctor Cannabis Doctor Supporter

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    If your PC only goes to 10 PSI I would go for 45-60 minutes.

    As far as the still air box goes you don't even need lysol really. You can actually just use straight water and mist the air and inside of the SAB. When you glaze the inside of a SAB like that you trap any of the floating particles in the water on the sides , bottom, and even under the lid,

    If you want you can lysol the air outside the SAB but even there you don't need it because of the same principle.

    You mist, water particulate gets trapped in the water droplets, and they land sticking them to the ground until the water evaporates.

    This is a lot healthier than using lysol.

    You can even use 10% bleach in the water if it makes you feel better but even that isn't necessary.

    Not that I am saying I have never done exactly what you did, lol.

    It is a paranoid game until you learn how to play it inside and out.
     
  15. nomendubium

    nomendubium scraping by, since '97 Expert Identifier

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    I feelya, prof. The lysol aought to do the same thing, but is propelled, thats why I use it. I realize I'm not killing anything, but just making it fall and stick to the liquid. Do you think the agar could have been that dirty? There are literally 100+ particles in each one. I've been using this sab successfuly for over a year
     
  16. Professor PinHead

    Professor PinHead Lost in the Tek.... Administrator Mushroom Doctor Cannabis Doctor Supporter

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    Like rogue said the time at that PSI wasn't long enough.

    If it was from you there would have been nowhere near that many inoculation points.

    Human error usually lands a patch or two of green or a pond of bactieria.

    PC error looks like that.

    There is only one way to learn though bro.

    Trail > error > Trial > Success

    I had a 12 PSI cooker when I was a beginner. They work, just require a longer cook.
     
  17. Jeff

    Jeff Well-Known Member Moderator Mushroom Doctor Supporter

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    Is it growing inside of the agar also or just on the surface? I couldn't tell from the pics
     
  18. nomendubium

    nomendubium scraping by, since '97 Expert Identifier

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    all of the white "patches" are on the surface, but shit loads of the flecks are all through it (look at the pic where I'm holding it up in front of the window) I pc'ed it in a pint jar with a solid lid. After I boiled it, and poured it into the jar, there was some dark brown sludge in the bottom of the pan=maybe I burned it?
     
  19. Jeff

    Jeff Well-Known Member Moderator Mushroom Doctor Supporter

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    If it is inside either your plates were contaminated or your agar. My bet is the agar (unless there was a hole in the bag of plates). Definitely need to pc longer at that pressure.
     
  20. Professor PinHead

    Professor PinHead Lost in the Tek.... Administrator Mushroom Doctor Cannabis Doctor Supporter

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    Go easy when you cook it. That "brown stuff" sounds an awful lot like caramelized sugar.

    Make sure to keep the flame down low if you are going to dissolve the agar flakes. Try starting with cold water and stay on it while the temp comes up. You really don't even need to cook it though. All you need to do is make sure it is well mixed. I pretty much pour mine into the jar as soon as it breaks a boil.

    This is because it will cook in the PC anyway.
     
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