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 JetBlackNinja
Pimp-o-licious
Registered: 11/26/03
Posts: 495
Loc: $%^&
Last seen: 19 years, 5 months
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 The secret of Agar 
#2184933 - 12/17/03 09:22 AM (20 years, 4 months ago) |
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So get this. I found a place I can order sterile pre-poured Malt-Agar in pretrie dishes. "Just Add Spore" type shit.
How hard is agar to use in these G2G methods you guys talk about, better yet, what is the best thing to do to get from colonized agar, to fruit by the foot?
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ignuF
Monster Mycologist
Registered: 08/04/02
Posts: 172
Loc: Wonderland
Last seen: 7 years, 4 months
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It is best to streak the plate (add spores), use a really sharp knife to cut abount 1.5in. off the end of a roll of cling wrap (use this to wrap around the outside edge of petri to seal it and hold it together for easy handling), incubate at 80-85F, then when it is colonized flame sterilize a scalple, cut a small wedge (about 1/4in x 1/4in), transfer to a ready made steril jar of grain spawn (rye berries, millet, WBS....), and then incubate the grain. All of the transfers and such are best done in a glovebox or flowhood  after a couple days you should see the culture colonizing the grain and when you get a patch about 1.5in - 2in diameter then you shake the grain and wait another 3-4 days to shake again. Then you should be near 100% colonization Ready to spawn to your straw, dung, or straw and dung mix.
Here is a good place to get your prepoured PDA plates. These people are great and I have used them many times in the past  Prepoured PDA petri plates
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All the above information is for microscopic research only!
Life is short so make a dent in the world so people can tell you were here!
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stara
addict
Registered: 05/14/03
Posts: 473
Last seen: 8 years, 4 months
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I use Sda agar do you know what's that?And I made an agar of spore syringe two days ago.Do you think it makes a differenc hydrated spore in a spor syringe or is it better to use dry spor by working with agar?Peace..
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 YidakiMan
Stranger
 
Registered: 09/28/02
Posts: 2,023
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Re: The secret of Agar [Re: ignuF]
#2185240 - 12/17/03 12:28 PM (20 years, 4 months ago) |
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It is actually not best to streak spores. The end result it a huge mess of dozens of substrains. If you seek a pure substrain, bettter to use the technique outlined by Micheal McCaw in TMC.
Sterilize three screw type vials with 9ml of distilled water. Let it cool. Use your inoculation loop to scrape the print gently. Dip the loop into the sterilized distilled water to release the spores. Now take the other two vials. Using a sterile pipette or syringe, take 1ml from the spore dilution into a different vial. Shake the new dilution to evenly spread spores. Now use a different sterile pipette or syringe to take 1ml from the new dilution into your final prepared vial. This is a 1:100 dilution and will only give you 2-5 colonies per plate. Much easier to sort out than standard streaking procedure.
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JetBlackNinja
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Registered: 11/26/03
Posts: 495
Loc: $%^&
Last seen: 19 years, 5 months
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Re: The secret of Agar [Re: YidakiMan]
#2185300 - 12/17/03 12:59 PM (20 years, 4 months ago) |
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Quote:
YidakiMan said:
It is actually not best to streak spores. The end result it a huge mess of dozens of substrains. If you seek a pure substrain, bettter to use the technique outlined by Micheal McCaw in TMC.
Sterilize three screw type vials with 9ml of distilled water. Let it cool. Use your inoculation loop to scrape the print gently. Dip the loop into the sterilized distilled water to release the spores. Now take the other two vials. Using a sterile pipette or syringe, take 1ml from the spore dilution into a different vial. Shake the new dilution to evenly spread spores. Now use a different sterile pipette or syringe to take 1ml from the new dilution into your final prepared vial. This is a 1:100 dilution and will only give you 2-5 colonies per plate. Much easier to sort out than standard streaking procedure.
You lost me 100%.
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automan
blasted chipmunk
Registered: 09/18/03
Posts: 8,272
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i think it is best to Not put spores on agar unless you are try to clean up a dirty print. grow how you normally would and when you find a nice big mushy that you like the looks of, clone it. you do this by tearing the mushy in half from the base (it splits easily) and cut a sliver from the inside, close to the base. then just drop that on the agar and it will start to grow. you will notice that the mycelia will start to produce both cottony and rhizomorphic (like the veins on an oak leaf). not only will you get those 2 types of mycelia, but you will get different sectors of each (for the rhizomorphic, it will look like feathers sitting side by side.....these are different substrains :] ). you transfer each sector (substrain) to its own dish and repeat the process untill you have only one type of mycelia growing in the petri dish. what i do at this point is pour some more dishes and transfer each substrain to anothr dish, that way i can put these dishes in the fridge to be able to use at a later date.... you'll see why in a minute.
now, each dish you have is a substrain that you can grow out to see if you like it. be sure to label everything well, that way you'll be able to keep up with how each substrain you are growing is doing. after the grow is over, you will have a good idea of which substrains were prolific fruiters, which were big is size, and which had a little of all the qualities that You are looking for.
now all you do is go back to the samples in your fridge and keep the ones that you liked. when you are ready for another grow, just take a little mycelia from that master dish and transfer it to another dish. as long as you have that master dish in the fridge, you will always have access to that original substrain that you isolated.
hope that helps some. feel free to ask for elaboration on parts that might not have been so clear.
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No, no, you're not thinking, you're just being logical. ~ Niels Bohr
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JetBlackNinja
Pimp-o-licious
Registered: 11/26/03
Posts: 495
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Last seen: 19 years, 5 months
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Re: The secret of Agar [Re: automan]
#2185364 - 12/17/03 01:34 PM (20 years, 4 months ago) |
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Ok, that I understand just fine, problem is, that sh1t doesn't matter to me at this stage. At this stage I want to get going and get good. Then I will worry about minute details such as "which substrain I like best". Right now I just want mushrooms.
I did the BRF's for awhile, now I am moving onto grains. I just want to see some success from these early grain trials. N o need for the advanced mycology theroy course.
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automan
blasted chipmunk
Registered: 09/18/03
Posts: 8,272
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ok..... i usually split 1 colonized dish to 3 pint jars.
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No, no, you're not thinking, you're just being logical. ~ Niels Bohr
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JetBlackNinja
Pimp-o-licious
Registered: 11/26/03
Posts: 495
Loc: $%^&
Last seen: 19 years, 5 months
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Re: The secret of Agar [Re: automan]
#2185414 - 12/17/03 01:52 PM (20 years, 4 months ago) |
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sweet, I use QT jars at the moment, so maybe 1 plate to 1qt jar?
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automan
blasted chipmunk
Registered: 09/18/03
Posts: 8,272
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no, 1:3 would still be fine.
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No, no, you're not thinking, you're just being logical. ~ Niels Bohr
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JetBlackNinja
Pimp-o-licious
Registered: 11/26/03
Posts: 495
Loc: $%^&
Last seen: 19 years, 5 months
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Re: The secret of Agar [Re: automan]
#2185443 - 12/17/03 01:59 PM (20 years, 4 months ago) |
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you the man.
Now my understanding is that getting Agar into WBS without contams is 1 of the hardest shroomin releated things to do, I am not spending 2G on a flowhood, but I will build a large glovebox if I can be assured the plans I follow are optimal.
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llamaboy
the weasel thatsnagged the bee
Registered: 11/08/03
Posts: 563
Loc: Portland PNW
Last seen: 14 years, 6 months
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use your oven...so i've heard, it's the best way...turn it on to warm, pull out the top rack and do everything on that...the warm air russhing up keeps contams from falling on to your open spawn...i guess this is related to a laminar flow hood...but that's just soemthing that i read... no idea if that's what it really is like.
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