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OfflineJericho_Button
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My 'SnackPlate' Method...
    #26350669 - 11/28/19 02:12 PM (4 years, 2 months ago)

So...

I'm pretty new to the game and I have an agar method I came up with that I'm sure most people would probably not want to bother with. It's a bit over the top but I wanted to make absolutely certain that I would be successful, and that it would limit the chance of mess.

My recipe is unimportant as I've been using someone else's, and I guess that the mixture depends on what you're growing. I decided to jump in the deep end with Pan Cyans, but I also have some Cubes running alongside them.

I don't have the resources or space to be doing large batches of agar all at once, and I only have a SAB so I wanted to do pre-poured plates. I know there are teks out there for this type of thing, and I had tried to cook some agar in a mini mason jar, but I found the depth of the vessel made it an absolute nightmare trying to make transfers (my hands were going over the mouth of the jar and I couldn't see shit!).

What I decided to do was to buy a small collection of borosillicate petri dishes (I have 12 x 90mm dishes), which are pretty pricey, but I didn't buy them all at once so it didn't seem so bad :tongue2:

I cook the agar, pour it in the dishes (somewhere between 20ml - 30ml per plate), then I let it cool and harden before putting the plates into self adhesive sandwich bags:



I get as much air out of the bags as possible whilst sealing them, then I twist the ends up and fasten them further; I usually use elastic bands but tonight I decided to use some zipties that I had lying around.



Then in to the PC:



When the bags come out they're usually vacuum sealed which I think is pretty awesome. I have had some problems with bags splitting due to not getting enough air out, using weak elastic bands, and cooling them down too quickly causing rapid expansion of the remaining air in the bags. I've also had some spillage a few times but I rectified that pretty quick by sorting out my trivet, the plates were still usable too. One final thing to look out for is: don't stack them too close to the lid of the cooker or one of the valves may get blocked...

I'm doing 45 mintues with my plates as my cheap PC only has a rocker, and I can't find any information pertaining to its operating pressure, so I thought I'd be safe. I've not had any issues so far, very little contamination. I'm having a problem with slow growth, but I have a feeling I've been using a hyper-nutritious recipe for transfer plates. The batch I'm cooking tonight is extra low nute so we'll see how it goes...


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Edited by Jericho_Button (11/29/19 04:43 AM)


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OfflineeLeSDenes
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Re: My 'SnackPlate' Tek... [Re: Jericho_Button]
    #26350685 - 11/28/19 02:25 PM (4 years, 2 months ago)

Nice 'tek'. If you want to make more, you can use spawn bags too, they are tall and could fit even 20 plates :smile:


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OfflineJericho_Button
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Re: My 'SnackPlate' Tek... [Re: eLeSDenes]
    #26350691 - 11/28/19 02:28 PM (4 years, 2 months ago)

That's not a bad idea! I'm on doing: print > agar > grain jars > substrate at the moment. I want to get a bit more experienced with these methods before I try spawn bags. I've also forked out a ton on gear so I don't wanna have to get a bag sealer right now. They do seem pretty convenient and I have successfully made liquid culture before so I feel I could get on with it.


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OfflineeLeSDenes
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Re: My 'SnackPlate' Tek... [Re: Jericho_Button]
    #26350768 - 11/28/19 03:34 PM (4 years, 2 months ago)

What I meant that you can put the petri dishes in the spawn bag which is bigger than a sandwich bag, and you don't need an impulse sealer just use a cable tie :smile:


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Offlinejbgtaa
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Re: My 'SnackPlate' Tek... [Re: eLeSDenes]
    #26350787 - 11/28/19 03:44 PM (4 years, 2 months ago)

So this...tek..? Is just you telling us how to PC glass petries? You don’t need to do it in sandwich bags, and you don’t need to sterilize the Petri dish itself. It’s cheaper to buy petries in bulk and pour them in the still air box, considering price per unit. 80 dollars for 500 petries is much cheaper than 80 dollars for 20. Plastic Petris come pre-sterilized and I’ve never had one contaminate in its own.


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OfflineJericho_Button
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Re: My 'SnackPlate' Tek... [Re: jbgtaa]
    #26350819 - 11/28/19 04:10 PM (4 years, 2 months ago)

Like I said, I know it's over the top, and I know I don't need to bag them. I chose to do it to prevent getting agar in the cooker if they boil over and prevent excess moisture getting inside the plates. The vaccum seal created means the plates can be stored pretty long term, and it also cuts the risk of contamination at the stage of pouring, and considering the fact that I only work with small batches of plates at a time it seemed like a logical choice.

I know it's needlessly expensive but I like it so I thought I'd share it, it's something a little bit different... It's not like I'm hyping it as 'the next big thing that everyone should be doing' or anything, I just wanted to be social.


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OfflineJericho_Button
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Re: My 'SnackPlate' Tek... [Re: eLeSDenes]
    #26350821 - 11/28/19 04:12 PM (4 years, 2 months ago)

Yeah I thought  of that after I'd posted haha! I might try it sooner rather than later, I think it would be a good idea as a spawn bag would be less disposable than the sandwich bags, and as you mentioned I could do larger batches. I have a second cooker that is larger so if I had the dishes for it I could do loads!


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Offlinejbgtaa
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Re: My 'SnackPlate' Tek... [Re: Jericho_Button]
    #26351264 - 11/28/19 11:17 PM (4 years, 2 months ago)

BUt what is your beef with pouring into sterilized plates?

When you buy a case of petris, they come in sleeves of 20, completely sterilized by radiation. This means you only have to sterilize the agar.

This lead sme to ask: whats the point of all this? Pastywhyte has pioneered the pastyplate revolution, and this "tek" youve posted is the same thing but using glass petris instead of ziplok rounds.

This is not a tek. And its honestly not practical for anyone who ACTUALLY cultures and isolates on agar. I couldnt use glass petris even if i wanted to, just because of the sheer amount of transfers i do. That alone means i have to make as mush of the process as disposable as i can.

If the bottom line is cost, then petris and a pyrex media bottle is a better option than what you have described in this thread. I have spent less than 100 dollars on agar suplies in total and still have enough to last me a year. You can get a case of petri's for 70 bucks, and agar and nutrients for another 20, and those materials will last you AT LEAST a year.

If the bottom line is sterility, well, that's on you. I have poured 100 petris at a time and not one of them contaminated. That alone should tell you that the entire process you use is unecessarily time wasting and expensive :shrug: sorry


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Re: My 'SnackPlate' Tek... [Re: jbgtaa] * 1
    #26351321 - 11/29/19 12:49 AM (4 years, 2 months ago)

Its alot of work being environmentally friendly but anything we can do to reduce single use plastic waste is definitely a good thing. It's a serious problem. I pour petris but sparingly my go to is the little anchor half pint regular mouth jars. I have like 3 maybe 4 dozen of them I keep in rotation.
They are cheap like 7 bucks for a dozen. Work good enough but they suck for visability for this reason I'm thinking about buying some glass petris to supplement my no pours.
Remember change isn't easy but it's 100% necessary.
Or some shit like that.:shrug:


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OfflineJericho_Button
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Re: My 'SnackPlate' Tek... [Re: jbgtaa]
    #26351398 - 11/29/19 03:06 AM (4 years, 2 months ago)

I spent some time working on a mushroom farm in Peru. When I was there the guy used a big 1 litre glass bottle to cook his agar in the cooker. If my memory serves me correctly it was just like the bottle used in the agar segment of RR's 'Let's Grow Mushrooms' DVD. My cooker is quite small and couldn't fit a bottle like that in, I have a larger one but the bottle would have to lie down which would cause it to leak everywhere. At the time it hadn't occurred to me that I could just use another vessel. I know it seems obvious, but sometimes I just don't think like that, it frustrates me.

I don't have any beef with pouring after cooking, and now I have a bit more experience with agar I will likely try it at some point as I do wish to start expanding my practise a bit and also working with many cultures simultaneously. As per usual, I was being overly cautious, but where's the harm in that?

I don't really see how it's a waste of time, the time taken to prepare the mixture, pour it in the plates then cook surely wouldn't take any more time to prepare the mixture, pour in a bottle, cook, then pour into plates... I would say that the time taken to get the pre-poured dishes into the baggies is probably not much longer than the time taken to get the prepared agar mixture into the bottle.

As far as waste is concerned, this is certainly something that I take into consideration. These baggies aren't reusable after cooking as the self seal adhesive strips become fused during the process meaning that I have to tear the bags open. Plastic petri dishes aren't reusable (or at least shouldn't be!), but consist of far more plastic than the baggies, and I can cook a maximum of 4 plates per bag. Zip ties and elastic bands can be done away with by using twist tag things, whatever they're called, which would certainly get a lot of use per item, and the same could be said for spawn bags if I choose to try them out (maybe those 'cook in the bag' oven things would also work?).

I appreciate that this method isn't practical, particularly when working with large quantities of plates as you do, but at this point I've not worked with more than 9 plates at a time. I also understand that this low number will bring difficulties with actually achieving what the transfer process is used for due to limiting my sample rate, but it's by no means impossible, and I'm still a novice. Again, 'like I said': it works for me.

The only beef I see is yours with my method. I don't mind criticism, and I feel I indicate clearly in my original post that I understand many people probably would have criticisms:

Quote:

Jericho_Button said:
I have an agar tek I came up with that I'm sure most people would probably not want to bother with. It's a bit over the top




The problem is, every time I offer you my reasoning for my decisions you don't seem to want to take it into account. Also, I'm not sure if I actually indicated that cost was necessarily a factor, especially not 'the bottom line'; in my original post I stated my recognition of this method of being 'a bit pricey' due to the glass dishes.

I would say sterility was a major factor when it came to my decisions, but your response seems to indicate that you have the impression that I've been having trouble with it. This isn't true. I also stated in my original post that I've had very little contamination, and the plates that do contaminate are usually ones that I had applied the parafilm to incorrectly. I think I'm actually excessive with my clean practice but I think continuing this is a sure way to ensure continued success.

One thing I would like to ask is: what exactly do you mean by 'this is not a tek'? Because I noticed that eLesDenes also seemed to disagree with my use of the term. As far as I understand tek basically means technique, and this IS a technique, whether practical or not. If I've misunderstood what 'tek' actually means please let me know.


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OfflineeLeSDenes
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Re: My 'SnackPlate' Tek... [Re: Jericho_Button]
    #26351422 - 11/29/19 03:48 AM (4 years, 2 months ago)

Dude I think your tek is valid and I only put them in quotation marks cause it was short (but its still a tek :laugh: ). I was always wondering how people have success with glass petris and you just showed a method which works for you. I think that's awesome. I particularly liked the vacuum bit with the bags which is the real game changer, especially if you use larger bags.
I work with plastic petris and go through a few sleeves in a very short amount of time and I am sick of dumping so much plastic after they are done. RR made a point a few years ago that washing the glass plates and sterilising is less environmentally friendly than just use the plastic ones, but I disagree. I think that pouring to plates and then sterilising the whole thing consumes the same amount of energy as sterilising a media bottle and pour later.


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Offlinejbgtaa
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Re: My 'SnackPlate' Tek... [Re: Jericho_Button]
    #26351423 - 11/29/19 03:50 AM (4 years, 2 months ago)

Brother you’re fine. I’m not mad and don’t need you to explain yourself to me.

All I’m saying is a TEK is something proven, peer reviewed, and usually disseminated by a credible source with a track record of credible information. What you are describing has been done by many and been proven to be a less superior route of culturing. Culturing has been simplified to plastic sterile petris and 500ml Pyrex media bottles (I’d be willing to bet my entire saving it will fit in you PC) and a scalpel and parafilm/ plastic wrap. No need to write a tek, if this did qualify as one.

If this post held something truly innovative, then I would call it a “write up”, but sadly it doesn’t.


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OfflineJericho_Button
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Re: My 'SnackPlate' Tek... [Re: jbgtaa]
    #26351447 - 11/29/19 04:40 AM (4 years, 2 months ago)

Oh I see, yeah that makes sense. I'll change the 'Tek' in the title to 'Method'.

With regards to the bottle, a half litre one probably would fit for sure, that's what I mean by 'using a different vessel'. In my head I had pictured the type of bottle I had seen used by others and for some reason it hadn't occurred to me to use a different bottle, however stupid that sounds haha!  :eek:

I haven't stumbled across a thread that is using this exact combination of materials, so as far as I'm aware it's unique, but I didn't expect it to be 'innovative', so apologising as though I would be offended by you stating that it isn't seems odd to me, but I can see how my use of the term 'tek' would lead you to believe that maybe I thought it was haha! Sorry about that. I've bought pre-poured plates in the past that came autoclaved in sealed bags. I do see what you mean when you say how the method of creating plates has been 'simplified' by trial and error of a large volume of people, but I don't necessarily agree that this method is more complicated. Sure, you could say that it's more expensive, and also excessive, but I had never poured plates when I first used this (I think I've only done 8 batches in total) and I just wanted some added security. As mentioned in my last post I will probably change up my methods soon enough now I have some confidence. I also don't think the impact different methods can have on waste should be disregarded.

One thing I would like to repeat is that my only intention for sharing this was to 'be social', I thought I made that clear. I haven't really read through the forum rules, so if sharing things like this is frowned upon, maybe I'll refrain from doing so in the future, but I can't see the harm in it, especially if I haven't made any claims to it's efficiency etc.


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Re: My 'SnackPlate' Tek... [Re: Jericho_Button]
    #26351486 - 11/29/19 05:43 AM (4 years, 2 months ago)

If those bags aren't melting then I would think your PC isn't cooking at the proper pressure...


Edited by SpunkyMonkey88 (11/29/19 05:43 AM)


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OfflineJericho_Button
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Re: My 'SnackPlate' Tek... [Re: SpunkyMonkey88]
    #26354970 - 12/01/19 04:41 AM (4 years, 2 months ago)

Yeah I think it runs at a lower pressure, hence the fact I extend my cooking times slightly.

I've had 0 contamination on my fresh agar, only 2/3 occurences on original plates after transfers, so I've introduced something during the process. Also, no grain contamination as of yet so I think it's serving it's purpose.

I wouldn't necessarily assume the plastic would melt, there's a braod range of different plastics, some are far more tolerant to high temps, and these bags seem to be one of those compounds. The zipties I used on this batch all broke, whereas the elastic bands suffer some deformation, but due to being flexible don't succumb to the pressure when the bags expand.

There were a further drawback I was painfully reminded of when I did my transfers with these plates the other night:

The dishes always come out of the bags wet with some condensation on. This has posed no problems other than when it comes to sealing the plates with parafilm. I use latex gloves and wet glass is slippery as fuck, so applying the parafilm is goddamn nightmare. On one occasion the plate slipped from my hand and landed agar side down on my belly. Needless to say a contaminant was introduced....

I had a higher nute mixture separated on one plate for spore germination, the bag with the plated in had fallen from atop the two stack os lower nute plates bellow, so when I removed it from the PC the agar had leaked inside the bag and solidified. Gutted.

On other thing to note: I've never had this happen before but one of my plates hasn't solidified after cooking. The only reasonable explanation seems to be that where the zipties broke, the bags came unsealed and the water line was above the lower level of the bottom dish, maybe excess water diluted the mixture? My transfer is still growing happily in there (for the time being), so it's kind of like LC in a petri haha!

I'm thinking of trying a post-pour method similar to this, but I'm hoping if I put the dishes into the bags empty and cook them, they will come out dry, and it will probably be easier to do a larger batch without any problems, I have a small 1/4 pint mason jar that would be perfect.


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