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InvisiblePastywhyteMDiscord
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Pastywhyte's Easy Agar Tek * 198
    #19208976 - 11/30/13 03:10 PM (10 years, 3 months ago)

Easy Agar Tek.

I'm starting to see an abundance of people out there who want to get into agar but are intimidated by the thought of pouring petris in a Still Air Box (SAB). There are others who are confident in their technique but don't like the idea of ordering lab equipment off the net to their house. Others still, dislike the idea of waste and having suspicious shit in their trash.

To get around this people are turning to no pour agar teks, and there are many good ones out there, they served the basis of how I myself got started with agar. But most of them have drawbacks. Jars are costly, half pints are too deep to work in with ease, fancy jelly jars are unwieldy to handle and quite expensive. The screwtop ziplocks would be a good alternative except they are large and take up a lot of space in the Pressure Cooker (PC) and fridge.

Enter "Easy Agar". I am here to say that none of the above mentioned drawbacks should dissuade anyone from getting into agar. In my opinion, agar does not need to be difficult, costly, or create a lot of suspicious waste. Many of the pro's and TC's will most likely flame this tek but, its not being written for them. This is for the new cultivator who wants to experience the benefits of agar without the cost and having to order stuff off the net. Let me say that I do use pour agar and proper petris, but for the lions share of my culture work this tek has served me well for 100's of plates.

My PP5 glad mini rounds are much smaller than 1/2 pints and screw top PP5, easier to manipulate in the SAB, and can even be made with a ghetto ass PC that you might have got as a wedding present and never needed or used (like the one I am using for this tek :lol:)

You will need:
-1 pack of Glad mini rounds
-micro pore tape
-a drill with a 1/4" bit or very hot nail
-paper towels
-Foil
-PC
-Bar of agar agar
-Potato flakes
-Karo or Honey
-Coffee grinder
-And of course a PC, even a shitty ass one will work





First we will prep the lids. They need a hole in them, to provide a bit of Gas Exchance (GE) as well as to prevent a vacuum when its PCing from crushing in the mini round container. You will need to drill a small hole in each lid. You can also just heat a paper clip or nail and use it to melt a hole in the lid, this is pretty quick. The hole does not need to be large, a 1/4" is best. Next cover the hole with a small square or two of micro pore tape. If I am planning to keep the plate around for a long time (more than a month) I would suggest two layers. Ok we're done, these are ready to fill :thumbup:





Now we need to prep our agar. Take the bar of agar agar and cut it into small chunks, then place them into the coffee grinder. Grind them until they are pretty fine. Don't worry about making them into powder, just get them pretty uniform. I have even used scissors to get er done, though a coffee grinder is easier. I prefer to grind up my bars and then just store the flakes for when I need them. 2 bars will go a long way. Feel free to use any other agars if you want, powdered agar packets works excellent, just make sure there is no added sugar.





I then spoon 2 heaping tablespoons into the pot. I rarely weigh it out, if I want stiffer agar I use a little more, if I want softer I use a little less. If you want to weigh it, for this amount of agar you will want about 2-2.5 grams depending on how soft or firm you want it to be. If your using powdered agar and don't have a scale, a level tsp (5ml) is roughly 2.5 grams. Remember that no matter how much you want to mix up, you want a ratio of 20-25 grams of agar to 1 liter of water. If you have issues with the mix being too stiff while mixing then adjust the ratios for less agar. Some food grade agars will work out to be a little stiffer than others, some experimentation may be required.

If all that is a bit confusing the just remember 2% is roughly what you are aiming for.

Add 2 teaspoons (10 ml) of potato flakes, and 1/2 cup (120 ml) of water (tap is fine). Then I turn the heat on low on the stove a stir the mixture until its mostly dissolved. At this point I will add a drop of Karo or Honey. Stir it all up well. It does not matter if there are still a few pieces of agar still not dissolved, just get most of it. I like to use either colored agar (some places have this) or else just add a few drops of food coloring to the mix. This will help you see growth real quick, and the contams are easily spotted as well. There will be nowhere for them to hide :lol: If you can only get regular agar, I like to use green food coloring to make the contams stand out best, but any dark color (red, blue, purple) will work as well :thumbup:





Finally I spoon in the agar mix into my glad mini rounds. I like to just give them a wipe with a damp paper towel first to clear out any dust or debris. No need to be spraying them with alcohol or nothing. Place the lids on the top and close. Finally I like to fold up a square of paper towel and place on top. Condensation can be a bit of an issue and you don't want to get the micro pore tape too wet. The paper towel helps with this. Here is the agar in the PP5's and how I do the paper towel.





Now we wrap them in foil before we place in the PC. I like to tear off a piece that is rather square, this will be a good size. Have the tab on the lid directed towards one of the corners of the foil, this helps get enough around and prevents tearing. To help get the bottoms flat I like to grind them down a little on the stovetop, this will help them sit straight in the PC. Place rings in the PC and a piece of folded up foil down if you need more stability. Stack carefully in the PC.





It was suggested by a poster (sorry I can't remember which one) that the hole could be drilled through the side and eliminate the need for the foil and towel. I have tried it and it does work. The tape gets quite wet and must be allowed to dry but it does work fairly well. I do miss not having the towel to write on and it's hard to not touch the tape sometimes so care should be exercised when handling. This would be a good option for those who don't want to use the foil.





Now you just gotta PC them for 40-45 mins. Done.  I like to leave them in the PC overnight to cool, this helps keep the condensation down. Then I when remove them they go straight into my SAB. If you want to keep em around for a while they can be stored safely in the fridge, just don't unwrap em. Remove the foil from them in the SAB and if the tops seem wet, I let them dry in the SAB for a few hours. Leave one of the arm holes open if you do this with a tyvek sleeve stuffed in the hole. Like I said before however, if they feel wet, adjust the way you bring the pressure up in the PC.

Now you have lots of cheap, easy to work with plates that don't need to be ordered and can be put together with stuff you can find around town. You can now safely inoculate any media like grains or LC without the issue of invisible contams. You can also store good cultures for a long time and grow them many times before senescence hits. You can also ward this off by changing your media, this tek ends up making PDA (potato dextrose agar) but it will work just as well with MEA (malt extract agar), PDYA (potato dextrose yeast agar), Dog food agar, grain soak water agar, etc. So don't be afraid to mix it up. Hopefully this lets people dip their toes in the agar pool, trust me its well worth it :awesomenod:



:scarface:

Edit; as more people come on board with this, I hear feedback on ways to use it, substitutions, etc. Here are some relevant links to assist users. I will attempt to update periodically.

The Tiger Drop

Inoculation, transfers, grain inoculation in SAB

Tic Tac Toe (Tiger Drop 2.0)

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OfflineTheApprentice
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Re: Pasty Agar Tek [Re: Pastywhyte] * 3
    #19209034 - 11/30/13 03:29 PM (10 years, 3 months ago)

I love the idea of food coloring!

great write up.

I started my agar adventure on brf paste in a glad mini round lol.

now I love agar!


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Re: Pasty Agar Tek [Re: TheApprentice] * 6
    #19209950 - 11/30/13 07:51 PM (10 years, 3 months ago)

awesome TEK!  I got some personal PM tutoring from pasty about this last week. I have transfers growing on them right now, will post pics as soon as the growth is big enough to see (just a tiny wedge now).  I dunno why, I hate pouring agar, I love doing transfers and stuff, but I dread the pour like I dread going to work.  With this its all ready to go after it cools in the PC!  I can't help but think this will encourage a lot more beginners to start agar.

YO THAT SHIT SO PASTY YO!  (sorry I work overnight at a convenience store in the ghetto, the lingo rubs off occasionally....)

:rofl:


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Re: Pasty Agar Tek [Re: blindingleaf] * 8
    #19210133 - 11/30/13 08:48 PM (10 years, 3 months ago)

Is this the testimonials section?  I spent a lot of time and effort trying to fuck these up and ended up with a usable agar plate anyway. Props to Pasty for making this excellent tek.


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InvisiblePastywhyteMDiscord
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Re: Pasty Agar Tek [Re: Psilicon] * 2
    #19210150 - 11/30/13 08:54 PM (10 years, 3 months ago)

Quote:

TheApprentice said:
I love the idea of food coloring!




Me too, I can't believe more people don't do it :super:

Quote:

blindingleaf said:
I can't help but think this will encourage a lot more beginners to start agar.




I sure hope so :awesomenod:

Quote:

van der griegen said:
I spent a lot of time and effort trying to fuck these up and ended up with a usable agar plate anyway.




Loving it :super:

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Re: Pasty Agar Tek [Re: Pastywhyte] * 2
    #19214047 - 12/01/13 06:14 PM (10 years, 3 months ago)

i like this tek. :thumbup:


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Re: Pasty Agar Tek [Re: silverstem] * 2
    #19214066 - 12/01/13 06:18 PM (10 years, 3 months ago)

:rockon:

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Invisibleteknix
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Re: Pasty Agar Tek [Re: Pastywhyte] * 3
    #19214089 - 12/01/13 06:22 PM (10 years, 3 months ago)

It seems better to just use a jar if you wanna no pour.


"Study: Most Plastics Leach Hormone-Like Chemicals."

Study: Most Plastics Leach Hormone-Like Chemicals. Makers of water bottles, including Camelback, now sell products that don't contain BPA, a chemical that can mimic the sex hormone estrogen. But a new study says that even if they don't contain BPA, most plastic products release estrogenic chemicals. Mar 2, 2011

http://www.npr.org/2011/03/02/134196209/study-most-plastics-leach-hormone-like-chemicals

"As scientists become more aware of the chemicals that leech out of plastics and make their way into our bodies, they are uncovering a variety of health issues that result. This includes cancer, reproductive issues, immune system suppression and problems with childhood development just to name a few."

http://www.onegreenplanet.org/environment/plastic-toxins-are-leaching-into-your-food-and-water/

Another thing to consider is the outgassing of most plastics;

"Outgassing can be significant if it collects in a closed environment where air is stagnant or recirculated."

https://en.wikipedia.org/wiki/Outgassing

"Most plastics are made by a process called "free-radical polymerisation" in which small molecules are joined together into long chains. Although the small molecules (things like styrene and vinyl chloride) are highly toxic, the long molecules made by joining hundreds or thousands of them together (polystyrene and polyvinylchloride (PVC)) are non-toxic. At high enough temperatures, however, the free-radical polymerisation reaction goes in the opposite direction; instead of small molecules joining together to make big ones, the big ones break down to give off the poisonous small ones."

http://www.madsci.org/posts/archives/2001-01/978994422.Ch.r.html

Edited by teknix (10/03/17 10:51 PM)

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Invisibleteknix
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Re: Pasty Agar Tek [Re: teknix] * 2
    #19214149 - 12/01/13 06:35 PM (10 years, 3 months ago)

It also seems like it might be a bit difficult to observe the growth through the lid and detect any contaminates readily and early on.

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InvisiblePastywhyteMDiscord
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Re: Pasty Agar Tek [Re: teknix] * 3
    #19214182 - 12/01/13 06:44 PM (10 years, 3 months ago)

Quote:

teknix said:
Isn't it cheap to by a petri dish?



Sure if you don't count $20 in shipping, that's a lot to some people.

Quote:

teknix said:
I think you might have a problem with settling of the nutrients at the bottom of the container.



Nope, that's why we heat it up to dissolve most of the nutes. This and many other no pour teks take advantage of this. Fact is that if the nutes were not soluble, LC wouldn't work either.

Quote:

teknix said:
Do you find that you have bubbles in the dishes after you're done?



Never happened yet.

Quote:

teknix said:
It also seems like it might be a bit difficult to observe the growth through the lid and detect any contaminates readily and early on.



Nope I look through the sides. Quite easy to spot satellite colonies and bacteria especially against the colored agar, which makes them stand out.

Quote:

teknix said:
It's much more efficient, imo, to just buy some premade PDA, at put it in one jar so you don't waste all the room in your PC.




Why not just do it in your oven? I'm not even going to debate this with you here. It states at the top of the thread that this is for people who don't want to be ordering shit online, or paying high prices for stuff they could do for pennies. I'm sure that you can appreciate people not wanting to order suspicious items online to their homes given your reluctance to post a single pic of anything you claim to have done.

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OfflineYuri.Pono
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Re: Pasty Agar Tek [Re: teknix] * 2
    #19214188 - 12/01/13 06:46 PM (10 years, 3 months ago)

how many times have you used this tek with them same type of glad jars. just wondering how many uses til the warp & are useless?


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InvisiblePastywhyteMDiscord
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Re: Pasty Agar Tek [Re: Yuri.Pono] * 2
    #19214209 - 12/01/13 06:51 PM (10 years, 3 months ago)

Quote:

Yuri.Pono said:
how many times have you used this tek with them same type of glad jars. just wondering how many uses til the warp & are useless?



I have done as many as 10 runs with the same containers. Sometimes I get a bad batch that don't last as long. I'd say an average of 5 runs would be fair. That works out to 40 plates for $4. Since I started with these I could not say how many times I have done these, but I am doing about 25-30 a month.

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Invisibleteknix
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Re: Pasty Agar Tek [Re: Pastywhyte] * 2
    #19214219 - 12/01/13 06:53 PM (10 years, 3 months ago)

Quote:

Pastywhyte said:
I'm not even going to debate this with you here. It states at the top of the




Good, I never said anything about it. . .


People charge you $20 for shipping? Free shipping if you order enough stuff from amazon. 5 bucks for 20 of them, how much do you pay for your little containers?

http://www.amazon.com/Karter-Scientific-Plastic-60x15mm-Sterile/dp/B005Z4QU5U/ref=sr_1_1?ie=UTF8&qid=1385949104&sr=8-1&keywords=petri+dish

These ones are twice as much, which is half the amount for the same price:

http://www.amazon.com/Sure-Fresh-Storage-Containers-10-ct/dp/B006USXTSG/ref=sr_1_4?ie=UTF8&qid=1385949169&sr=8-4&keywords=small+container

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Invisibleteknix
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Re: Pasty Agar Tek [Re: Pastywhyte] * 2
    #19214221 - 12/01/13 06:54 PM (10 years, 3 months ago)

Quote:

Pastywhyte said:
Quote:

Yuri.Pono said:
how many times have you used this tek with them same type of glad jars. just wondering how many uses til the warp & are useless?



I have done as many as 10 runs with the same containers. Sometimes I get a bad batch that don't last as long. I'd say an average of 5 runs would be fair. That works out to 40 plates for $4. Since I started with these I could not say how many times I have done these, but I am doing about 25-30 a month.




Where did you buy those at?

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Re: Pasty Agar Tek [Re: teknix] * 3
    #19214228 - 12/01/13 06:57 PM (10 years, 3 months ago)

The grocery store. $4 is the high price. A couple months ago they had them on sale for $2.50 and I loaded up. Might even be cheaper in the U.S. We seem to get hosed on prices for everything up here.

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Invisibleteknix
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Re: Pasty Agar Tek [Re: Pastywhyte] * 2
    #19214245 - 12/01/13 07:01 PM (10 years, 3 months ago)

What grocery store and For how many of them?

I guess if you can reuse them you might save a bit, but I hate cleaning plastic that has been melted with agar and nutes stuck in the sides.

I used to use pyrex because I thought it would be cheaper in the long run, but really it seems more of a hassle when I could do better things that clean up all the dishes, plus those would take up quite a bit more space. Space is money too.

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Re: Pasty Agar Tek [Re: teknix] * 2
    #19214308 - 12/01/13 07:16 PM (10 years, 3 months ago)

Nice tek and write up. Well documented  - with photos. Imagine that...

Thanks

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Re: Pasty Agar Tek [Re: budkatz] * 3
    #19214359 - 12/01/13 07:31 PM (10 years, 3 months ago)

I've seen them in every grocery store I have been in. They are not hard to find. Truth is they are not hard to clean. Soak em for 5 min in soapy water, then a quick wipe with a dishcloth. Done. I can wash up 30 in about 7 min including the 5 min soak time. Now it's even easier cause I just get my wife to do it :lol:

Quote:

budkatz said:
Nice tek and write up. Well documented  - with photos. Imagine that...

Thanks




Appreciate it man, good vibes :super:

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Re: Pasty Agar Tek [Re: Pastywhyte] * 2
    #19214426 - 12/01/13 07:48 PM (10 years, 3 months ago)

Nice, Pasty!

Quote:

Pastywhyte said:
Quote:

Yuri.Pono said:
how many times have you used this tek with them same type of glad jars. just wondering how many uses til the warp & are useless?



I have done as many as 10 runs with the same containers. Sometimes I get a bad batch that don't last as long. I'd say an average of 5 runs would be fair. That works out to 40 plates for $4. Since I started with these I could not say how many times I have done these, but I am doing about 25-30 a month.



Those should last you far more than 5 uses.
Mine are screw-top of course but I am quite sure that many of mine have had more than 50 runs in the cooker.  All of them have certainly had more than 10 runs.
Only visible wear is scratching from using scrubbies a couple times to clean them out.

Also I never get a "bad batch" of the screw-top ones, at least not with zip-locs or the most similar generics (the purple/blue lid kind they re-label for different marts)

I'll add, screw-top containers don't need lid modifications!


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Re: Pasty Agar Tek [Re: Violet] * 2
    #19214509 - 12/01/13 08:10 PM (10 years, 3 months ago)

Thanks Violet. A few things on your well founded comments.

Quote:

Violet said:
Those should last you far more than 5 uses.



For the most part they do, I was just trying to compensate for the occasional bad batch which I have run across, driving the average down. Fact is that I still have some of the very first ones I ever did and have probably gone well over 10 runs, just trying to be on the safe side of things.

Quote:

Violet said:
Mine are screw-top of course but I am quite sure that many of mine have had more than 50 runs in the cooker.  All of them have certainly had more than 10 runs.



No question. I have used your screw top ziplocks many times back when I was first experimenting with no pour well before I had an account here. My reasons for not using them is ergonomic rather than durability. I find dealing with the large size of the container and the screw lid cumbersome. I can hold the mini round and crack the lid with one hand allowing the other hand free to use my tools and exposing very little of the container to the air in the SAB. Plus I can fit many more mini rounds in my fridge which is a plus when you get over 50 cultures on the go.

Your ziplocks work perfectly fine for most purposes. I did successful agar with them before I ever saw your tek. It all comes down to personal preference. For me ergonomics and compact size trump durability and not needing to modify the lids :super:

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