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OfflineLtLurker
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Re: New guy. Trouble navigating [Re: ErnieErgot67]
    #27154008 - 01/17/21 08:30 PM (1 month, 9 days ago)

ditch the tin foil. it's not gonna do much.


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OfflineErnieErgot67
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Re: New guy. Trouble navigating [Re: LtLurker]
    #27154402 - 01/18/21 12:51 AM (1 month, 9 days ago)

Thanx. I did. I figured the extra piece of tape would be plenty. Plus now I’ll be able to see them easily and I won’t have to pick them up when I look at them every hour to check their progress!!  LOL ha ha just kidding but you know what I mean!  It’s the equivalent of… “are we there yet?!are we there yet!?“ x ten and on steroids.
Well as soon as this bottle cools down to the right temp I will be pouring my first agar plates! Wish me luck, say a prayer and cross your fingers that I don’t screw it up and that I don’t contaminate any of them. Looking forward to chatting with you again.


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Can’t we all just get along???
Progress not perfection.
Don’t judge others because their flaws are different than yours


Edited by ErnieErgot67 (01/18/21 12:54 AM)


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OfflineErnieErgot67
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Re: New guy. Trouble navigating [Re: ErnieErgot67]
    #27154496 - 01/18/21 02:31 AM (1 month, 9 days ago)

Well now I’m back to totally confused again. I thought the purpose of making these plates was to avoid contamination. Now after reading C 10 ‘s post on agar and inoculation tips I haven’t a clue what he’s talking about. What I got out of this is I am going to be deliberately inoculating my agar with my LC and my MSS and there is going to be bacteria in it and I’m going to contaminate my dish...DELIBERATELY!  I thought the point of making a guy was so I didn’t have bacteria. I didn’t think I was supposed to use this loop tool to colonize it!  I don’t even have this loopy tool thing that I’m supposed to make scratches with!  Now I’m right back where I started at another brick wall. Ah Fuck it! I’m just putting a drop of spore solution on one dish and a drop of LC  in another and see what happens!
That’s all I thought I was supposed to be doing.


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Can’t we all just get along???
Progress not perfection.
Don’t judge others because their flaws are different than yours


Edited by ErnieErgot67 (01/18/21 04:09 AM)


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OfflineGrinchGrower
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Re: New guy. Trouble navigating [Re: ErnieErgot67]
    #27154501 - 01/18/21 02:39 AM (1 month, 9 days ago)

That will work fine... the loop just helps you separate the spores/inoculant a little into smaller, more distinct colonies.
You will be making multiple transfers from one agar plate to another in order to attempt clearing up any bacteria from your initial germination.


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OfflineFungi Rogers
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Re: New guy. Trouble navigating [Re: ErnieErgot67]
    #27154582 - 01/18/21 04:15 AM (1 month, 9 days ago)

Quote:

ErnieErgot67 said:
Well now I’m back to totally confused again. I thought the purpose of making these plates was to avoid contamination. Now after reading C 10 ‘s post on agar and inoculation tips I haven’t a clue what he’s talking about. What I got out of this is I am going to be deliberately inoculating my agar with my LC and my MSS and there is going to be bacteria in it and I’m going to contaminate my dish...DELIBERATELY!  I thought the point of making a guy was so I didn’t have bacteria. I didn’t think I was supposed to use this loop tool to colonize it!  I don’t even have this loopy tool thing that I’m supposed to make scratches with!  Now I’m right back where I started at another brick wall. Ah Fuck it! I’m just putting a drop of spore solution on one dish and a drop of LC  in another and see what happens!
That’s all I thought I was supposed to be doing.




You confused me pretty good there bud. So agar is to check if it’s clean and to transfer away from contams


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Invisiblecoversall
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Re: New guy. Trouble navigating [Re: ErnieErgot67] * 1
    #27154651 - 01/18/21 05:36 AM (1 month, 8 days ago)

Quote:

ErnieErgot67 said:
Well now I’m back to totally confused again. I thought the purpose of making these plates was to avoid contamination. Now after reading C 10 ‘s post on agar and inoculation tips I haven’t a clue what he’s talking about. What I got out of this is I am going to be deliberately inoculating my agar with my LC and my MSS and there is going to be bacteria in it and I’m going to contaminate my dish...DELIBERATELY!  I thought the point of making a guy was so I didn’t have bacteria. I didn’t think I was supposed to use this loop tool to colonize it!  I don’t even have this loopy tool thing that I’m supposed to make scratches with!  Now I’m right back where I started at another brick wall. Ah Fuck it! I’m just putting a drop of spore solution on one dish and a drop of LC  in another and see what happens!
That’s all I thought I was supposed to be doing.




Agar is a predictable 2D surface to grow stuff on. It's a good way to start growth from potentially unclean sources as it allows for easier identification of myc and contam. Once both have been ID'd you can, with a high degree of confidence, move the growth you want to another clean plate leaving behind any contam. On the second plate you can then continue you to grow the sample, hopefully without any of the contam that appeared on the first plate.

The loop allows you to spread whatever it is you put on agar around, again in a controlled and predictable manner. This is commonly done in a certain pattern that allows you to a have a 'gradient' of concentration from point A to point B. With a higher concentration of 'stuff' at point A. As the streak moves towards point B the hope is that everything becomes spread out, making it easier to discern between contam and myc.

It's not so much that you are deliberatley contaminating your plates, but you are accpeting that the spore solution or spore print may well be contaminaed. So you are using the agar to seperate desired growth from contam, through streaking and transfers.

I'm typing this out as I'm supposed to be working! haha. Hope it helped!


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OfflineErnieErgot67
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Re: New guy. Trouble navigating [Re: coversall]
    #27156318 - 01/18/21 11:23 PM (1 month, 8 days ago)

Dude! Absolutely perfect. Yes it did help and it helped a lot. You explain to me exactly what was going on in that guys diagrams. I guess what I don’t understand is what the scratches are for and I don’t understand why they intersect and I don’t understand why they go around the outside of the dish.
Unfortunately my impatience got the best of me and of course I know best…a little Sarcasm there...and I went ahead and put a drop of the MSS on a plat, actually three plates And another drop on another plate of an LC. Hopefully I get from mycelium  going and I’ll be able to make up some of my own clean LCs and I will be able to inoculate some grain with agar and all the other things that we do with agar. Thanks for taking the time to explain that to me. Especially while you were at work. I very much appreciated that. ✌🏻💚🙏🏻😆🍄


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Can’t we all just get along???
Progress not perfection.
Don’t judge others because their flaws are different than yours


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Invisiblecoversall
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Re: New guy. Trouble navigating [Re: ErnieErgot67]
    #27156674 - 01/19/21 05:13 AM (1 month, 7 days ago)

The different streak patterns are just that, different patterns, all aiming to do the same thing. The simplest streak would be a single line across the plate, next maybe a zig-zag, which increases the length of the streak, spreading the 'stuff' out further. Beyond that you can create longer streaks, many different streaks, and so on, all aiming to spread things out in a predictable manner.

See a bunch of stuff growing where you didn't streak? Chance are it's not what you want!

At the end of the day you don't need to streak, but it helps, your plates may very well produce nice healthy myc! Only time will tell.

If you're anything like me you'll charge ahead, mixing a bunch of half read teks, have a mix of success and failure before finally settling down and actually following one tek to the tee and getting results! Hah! You just got to burn off some of the 'fresh out the gates' energy.

:rush:


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OfflineErnieErgot67
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Re: New guy. Trouble navigating [Re: LtLurker]
    #27157156 - 01/19/21 12:17 PM (1 month, 7 days ago)

Good morning LT,
    I wanted to ask you why it’s preferred to use the polyfill over the micropore tape? (Which I doubled up BTW.) With the the LC that I used, it was just over 24 hours and I already see mycelium starting to grow! I was and am amazed. I pray that all those jars of wheat berry colonize without any contaminants. So right now on the burner, I have,...14 BRF cakes, I have 6 quarts of inoculated  wheatberries on the burner, and I have 20 agar plates poured with four of them inoculated with 3 different types of strains or species. I’m not sure which word is the proper term...Strain? Species? I will guess the species is Psilocybe Cubensis and the strain is PE, APE, GT, etc...Which reminds me, have you ever heard of a strain with these initials because this is one of what was given to me...B.D.S.K.???
So I went from this super confused overwhelmed guy to having almost every iron in the fire involved with the mycology world...,well, not even close but I’m guessing you get the point. Haha. This is one of the reasons why I was so frustrated. Because I’m far from an idiot and of all the different types of knowledge in this world biology, Science, anatomy, physiology, health, nature, etc. etc. is my best subject. In high school I always had A’s in science and then I went to a tech college to get my RN, starting as a surgical tech, and I had highest grades in my class.(no exaggeration) It’s a long story why I never finished but I was 3 months away from graduation when my word crumbled. (Thru no fault BUT my own). On top of that, I’m a professional gardener by trade these days and for the first half of my life I was a chef and just in case you didn’t know cooking involves following recipes and directions. So you put all that education and trades together and you should have one almost perfect amature mycologist. Wouldn’t you agree? For the most part, humans are our own worst enemy. I have a tendency of over complicating things. And my pride can always trip me up. That little voice in my head telling me...what is wrong with you EE! you’re not an idiot! you should know how to do this!
Anyway, I really hope I’m well on my way. I know that this involves failure before success or at least failure involved with success just to word it on a more positive manner. Thanks again for your help and patience LT. I hope I can still count on you down the road because the whole Rison looks like a whole new unexplored world! Lol. One question before I end this book. It says to store the agar dishes upside down for condensation purposes. Can you store the already inoculated ones upside down for the same purpose or will that mess up the growth in the dish? I stack them on top of each other as I was pouring them but some of them still got condensation.


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Can’t we all just get along???
Progress not perfection.
Don’t judge others because their flaws are different than yours


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OfflineErnieErgot67
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Re: New guy. Trouble navigating /coversall and agar [Re: ErnieErgot67]
    #27157209 - 01/19/21 12:48 PM (1 month, 7 days ago)

Hey Brutha, well if you’re a dude! Lol. Sorry if you’re not.  Thanks again for the reply. I appreciate all the help I can get and I understand that people are busy.  Yes I would say I am like you and I will charge ahead. The only thing I did correctly on here when it came to listening to directions and suggestions was I started with the PF Tek. I wanted to just skip right over that and go to grain and bulk. However, if you go back through my post you will see all the people who attempted to help me while I was freaking out and so overwhelmed and they all suggested that I start there. Oh man, I was a mess when I first came on here. I was whining and bitching and moaning on how I couldn’t find anything on here and I couldn’t use the search engine… And I still can’t! LOL. I am extremely impatient when it comes to stuff like this and patience is one of the main things you need to be in mycology. It’s the fucking worst! You have to do all this work and you have to wait weeks to find out if you did it right. That is horrible! I mean that’s the worst! The best thing these guys did for me was not give me easy answers every time. They made me look them up and then ask questions. In doing so it let me down many rabbit holes and I read things that I didn’t really need at the time which eventually came full swing adding to pieces of the puzzle. I’m sure you understand. I was told to stay off of YouTube because the majority of their practices to say the least… Hacks! Well going on YouTube  help me out immensely because I was able to connect the dots by watching procedures. Trying to read directions on here without knowing even the vocabulary (especially the tons of abbreviations!!) made it hard to understand directions so by watching it helps to fill in the blanks. Also by watching YouTube it showed me what not to do. Instagram helped a lot because I can ask a question and get an answer immediately. Not for nothing but there are some pretty professional mycologists on Instagram. Because they show pictures of their labs and their results it’s pretty easy to distinguish between a hack and a pro. I noticed that you’ve only been out here for under a year. Have you been doing this a while and before you got on the Shroomery. I guess once I start playing with the agar I will try making the scratches and see what happens. Is there a different tool I can use to substitute for the one that they are talking about with the loop? Thanks again for all your help and sorry for the book I just wrote. I can be long-winded  Lol :thanx:


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Can’t we all just get along???
Progress not perfection.
Don’t judge others because their flaws are different than yours


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OfflineFungi Rogers
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Re: New guy. Trouble navigating /coversall and agar [Re: ErnieErgot67]
    #27157506 - 01/19/21 03:26 PM (1 month, 7 days ago)

I believe most people use polyfil over micropore due to it being cheaper and easier to find. Don’t quote me on that though :smile:

You don’t have to store your agar upside down I leave an extra poured Petri on top of my stack if I have condensation which just moves the condensation to the sides( try to pour the agar at perfect temperature to cut back on condensation/ before it starts getting thick again). Storing them upside down won’t affect the growth of the mycelium.
Uhm I’m not so sure on a substitution for an inoculation loop other then you can make your own? I just bought one for a couple dollars on Amazon lol.


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OfflineErnieErgot67
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Re: New guy. Trouble navigating /coversall and agar [Re: Fungi Rogers]
    #27161813 - 01/21/21 05:50 PM (1 month, 5 days ago)

I’m not sure where I’m supposed to post this in order for me to get some help with it so I just thought I would post it where I have been posting. I’m not sure if this is contaminant or if it is actual rust spots from water and the minerals in the vermiculite because I actually had this rust colored spot before The mycelium started growing but it never really grew but now it is and it’s growing with the mycelium. The mycelium  itself looks healthy. HELP. I pray that this is not a contaminant.  I would at least like to have a successful TF Tech and then get my failures as I drive more complicated stuff. I don’t wanna have a failure right off the bat!  These jars were inoculated on January 9. HELP! HELP!
PS.... don’t mind the black permanent marker drawn on the outside of the jars I was just marking where the last mycelium  growth was so I could gauge how quickly it was growing. I marked those two days ago so it’s growing pretty quickly.



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Can’t we all just get along???
Progress not perfection.
Don’t judge others because their flaws are different than yours


Edited by ErnieErgot67 (01/21/21 05:57 PM)


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OfflineGrinchGrower
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Re: New guy. Trouble navigating /coversall and agar [Re: ErnieErgot67]
    #27161896 - 01/21/21 06:25 PM (1 month, 5 days ago)

Looks like when I overcooked my jars, the bottom edge got a little toasty.
If it's changing size/shape though it's likely metabolites and/or contamination.


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OfflineErnieErgot67
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Re: New guy. Trouble navigating /coversall and agar [Re: GrinchGrower]
    #27161981 - 01/21/21 07:15 PM (1 month, 5 days ago)

I wish that’s what it was. Unfortunately it was not there from the beginning. But it was there before. And it looked like some of the vermiculite granules. That’s why I’m leaning towards oxidize minerals. Or metabolites like you said. I guess time will tell. If it’s a bacteria or a mold it will start taking over her my mycelium . Correct?


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Can’t we all just get along???
Progress not perfection.
Don’t judge others because their flaws are different than yours


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OfflineErnieErgot67
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Re: New guy. Trouble navigating /coversall and agar [Re: ErnieErgot67]
    #27162107 - 01/21/21 08:28 PM (1 month, 5 days ago)

I can’t believe I figured out how to do this but I found this post and I dragged it over to here.
Besides this post there was another one where it was suggested that while drilling the lids fine metal dust fibers and shrap could’ve been in the bottom of the jar and rusted. Plausible as well. However I’m going with the following link.
https://www.shroomery.org/forums/showflat.php/Number/16038384#16038384


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Can’t we all just get along???
Progress not perfection.
Don’t judge others because their flaws are different than yours


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OfflineFungi Rogers
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Re: New guy. Trouble navigating /coversall and agar [Re: ErnieErgot67]
    #27162364 - 01/21/21 10:45 PM (1 month, 5 days ago)

Hopefully all turns out well for you man! Just gotta power through you’ll definitely know if you start smelling anything weird or they start getting excessively different


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Re: New guy. Trouble navigating /coversall and agar [Re: Fungi Rogers]
    #27162462 - 01/21/21 11:35 PM (1 month, 5 days ago)

Well if they go south I have Plan B going on. I have 6 quart jars of wheat berry inoculated and it’s colonizing as we speak. The white stuff is spreading!

@whoever. I inoculated my agar almost a week ago and so far I am not seeing any bacterial growth. However the condensation is ridiculous and making it very challenging to see anything in detail. I have been keeping them in a temperature between 75 and 80°  Now that the liquid culture and spore liquid has dried or stop rolling around or wherever it went I turn them upside down to see if it would help with the conversation so I can get a better visual. Any advice? Thank you. Y’all have a great day God bless


Edited by ErnieErgot67 (01/22/21 07:27 AM)


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Invisiblecoversall
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Re: New guy. Trouble navigating /coversall and agar [Re: ErnieErgot67]
    #27163026 - 01/22/21 09:49 AM (1 month, 4 days ago)

The joy of the PF tek is that if the jars do look a little suspect you can top fruit them. That way you still get mushrooms and limit the spread of water contam you think may be present.


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Re: New guy. Trouble navigating /coversall and agar [Re: coversall]
    #27163800 - 01/22/21 05:25 PM (1 month, 4 days ago)

It's probably fine. I've had some verm that gave some bacteria looking red reactions to. Ride it out, worst case you top fruit bacterial jars.


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Re: New guy. Trouble navigating /coversall and agar [Re: LtLurker]
    #27164438 - 01/22/21 10:28 PM (1 month, 4 days ago)

Thank you. I guess that means spawning these to bulk is out of the question!? Unless it really is rust. Which I still haven’t ruled out.


Please have a look at these. Because of the condensation I don’t know what I’m looking at. I also don’t know what I’m looking at just because… I don’t know what I’m looking at! LOL. It’s the first time I’ve ever worked with agar. If I were to guess I would say everything growing in there is bacterial and not mycelium.



Edited by ErnieErgot67 (01/23/21 04:25 PM)


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Mushrooms, Mycology and Psychedelics >> Getting Started

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