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ar393
old timer
Registered: 01/26/00
Posts: 702
Loc: VT
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Got great growth on agar, now what
#348627 - 06/25/01 12:55 PM (22 years, 9 months ago) |
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My cat recently started playing with agar, and is very suprized how easy it is to work with, any ways after a few initial plates colonized, my cat took some sectors and transferred to a second set of plates. so now that the second set of plates are almost done (beautiful circular rhizo growth), she took a wedge from each and dropped it into a few rye jars. now i wonder if you could take the agar surface (since there is sooo much mycelium on it) and maybe blend it or some how get the myco into suspension so that instead of one large innoculateion point there could hundreds. and yes i understand that my cat could go via the honey water path, but agar is soo much easier to control contams , etc... I also worried that my cats jars will contam even though the rye was pre soaked, PCed at 15 psi for 45 minutes, then allowed to sit to see if anythign survived (2 weeks) and then PCed again for 30 minutes at 15 PSI before innoculation. the reason i did this for my cat is i fear that some rye grains on the opposite side of the agar (the side without myco) might deposit contams onthe agar, and the contams could take off. just my thoughts, any clarification or ideas or suggestions would be great. ar393
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Anno
Experimenter
Registered: 06/17/99
Posts: 24,166
Loc: my room
Last seen: 19 days, 13 hours
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Re: Got great growth on agar, now what [Re: ar393]
#348649 - 06/25/01 01:25 PM (22 years, 9 months ago) |
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You can transfer 1 healthy agar plate to a blender and sterile blender containing water blend it for a few moments. Then transfer the slurry to rye. Difficult to perform under non compleately sterile conditions though. If double steilizing( which is no need of), then after a few days, not weeks. I PC my rye 1 hour.
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ar393
old timer
Registered: 01/26/00
Posts: 702
Loc: VT
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Re: Got great growth on agar, now what [Re: Anno]
#348833 - 06/25/01 06:28 PM (22 years, 9 months ago) |
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yeah i was going to only leave it for two days, and i usually dont double steralize, but i had had some issues ,over 20 quarts contamed. and i was going to only wait a few days but my cat wasnt around.
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Azure
old hand
Registered: 12/31/98
Posts: 469
Loc: California, USA
Last seen: 21 years, 10 months
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Re: Got great growth on agar, now what [Re: ar393]
#348971 - 06/25/01 09:57 PM (22 years, 9 months ago) |
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You could take a tall half pint jar, fill it 1/4 filled with malt and water, sterilize at 15 psi for 20 minutes, and then under a flow hood or oven, transfer a sector into the malt solution. After a day or two, it will be fuzzy, and you swirl the solution around daily. When you have a bunch of milky mycelium settled on the bottom, you can suck that up and innoculate pf type jars that have birdseed instead of brf. That way, you'll have a fatter yield. The key is to not touch the lid when you open the jar, which can be somewhat tricky. But with practice, you'll find it's very easy to do.
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auto59009
enthusiast
Registered: 05/20/01
Posts: 376
Loc: Australia
Last seen: 22 years, 4 months
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Re: Got great growth on agar, now what [Re: Azure]
#349066 - 06/26/01 12:32 AM (22 years, 9 months ago) |
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hey ar383 i often make myco syringes in suspension of distilled water, as opposed to a spore syringe, simply because of the excellent rhizomorphic growth, so here is what i do; sterilize some babyfood jars by pressure cooking with lids off. then when done release steam, empty jars and quickly recap (i wear 3 layers of latex to avoid burns). now you have sterile jars. from here you create a sterile environment w/flowhood if possible. then simply wipe down a xacto blade with iso alcohol to sterilize. cut out a 1/2" x 1/2" square from the agar plate. drop this into the jar and add sterlised water from the syringe you intend to store the suspension in. shake the jars to remove strands of myc from agar cube. then draw up water and voila, a nice myco syringe i recently tried an update on this method by scraping myc off the agar (like a 2" square) and then adding that to the babyjar so there is a LOT of myc in there. yet even with a small cube, the visible strands are not indicative of all the mycellium. i find that the water becomes mycellium rich due to the shaking not matter how much myc is used. my guess is that even tiny groups of cells of myc would be distributed thru the syringe, and they are not visible to the naked eye. i have used this method many many time because it is so easy and the growth is fantastic. i originally saw the method posted at MMA and made some alterations to it. i don't own a flowhood or anything, just spray down an area with lysol and go from there. results are usually excellent and the contamination rate is quite low (in my experience anyway) hope that helps!
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MNmyc
enthusiast
Registered: 02/25/01
Posts: 175
Loc: MN
Last seen: 17 years, 20 days
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Re: Got great growth on agar, now what [Re: ar393]
#349067 - 06/26/01 12:33 AM (22 years, 9 months ago) |
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A small mouth jar accepts the blender parts and is pc'able.
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ar393
old timer
Registered: 01/26/00
Posts: 702
Loc: VT
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Re: Got great growth on agar, now what [Re: MNmyc]
#349180 - 06/26/01 06:57 AM (22 years, 9 months ago) |
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well my cat has dropped agar chunks into my jars and we will see what happens, its been about two days and im not seeing much, but it could take a little time for the myco to get used to the rye. since i didnt have rye flour or a grinder , to make the agar with rye flour for my cat. i also hope it wasnt too hot for the myco when my cat dropped it in the rye, but we shall see. worse come sto worse i lose it all and my cat has to do it again.
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