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Adustus
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Registered: 11/14/13
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Starting my mushrooms in some agar
#19253302 - 12/09/13 11:46 PM (10 years, 1 month ago) |
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So im going to use PastyWhytes Agar tek and use some live cultures I have.
I have a PC and made a SAB and have all the sterilizing stuff.
Anyone have any advice for a first timer using agar? And how to maybe make it go smoother? Im using jars instead of gladware.
I also was wondering if anyone had any ideas for a rack? My lid dont seem to wanna fit inside very well.
These are my only cultures so im really concerned with the inoculation process bringing contams. Any help is greatly appreciated!
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Forrester
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Re: Starting my mushrooms in some agar [Re: Adustus]
#19254542 - 12/10/13 09:45 AM (10 years, 1 month ago) |
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The biggest help I've found is once you have your SAB wiped down with alcohol and everything all set up in there and ready to go, leave it to sit for like an hour so any spores can have a chance to settle. I used to do 15 minutes because that's what I read, but I had a lot of contamination problems until I just started waiting longer. Also try to minimize the number of times you bring your hands in and out of the box.
If you're using ball jars like me, get your lid rings broken loose before you let it sit, because it's tough to break them loose in the box with your hands covered in alcohol and slippery. You wanna have everything placed right and a plan in your head of exactly what you're gonna do, do it quick and minimize the time you have the jars open.
Most people around here flame their scalpel blade between cultures, but I just use a new one so I'm not bringing my hands in and out of the box with hot blades and creating air currents. They're like $7 for a box of 100.
I'm no agar expert these are just a few things I've done to get much better results in my SAB. Also, I use the upside down box on top of moist towel routine that Notahacker420 shows here, this way any spores that fall stick to the towel and don't get stirred up again.
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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Amanita virosa
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Re: Starting my mushrooms in some agar [Re: Forrester]
#19255130 - 12/10/13 12:17 PM (10 years, 1 month ago) |
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Although jars work fine, consider investing 7 bucks in a sleeve of sterile plastic petri plates. MUCH easier to work with, esp. in an sab. take up less room in the fridge, less awkward to sub out of and easier to see contam. Otherwise, allowing your jars to cool with the lids slightly loose (flip the lids upside down, rubber side up) will allow excess moisture to escape. Otherwise, you may have some on the surface of the agar. Consider making some slants for long term storage of your cultures.
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Adustus
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Re: Starting my mushrooms in some agar [Re: Forrester]
#19256266 - 12/10/13 04:37 PM (10 years, 1 month ago) |
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Quote:
Forrester said: The biggest help I've found is once you have your SAB wiped down with alcohol and everything all set up in there and ready to go, leave it to sit for like an hour so any spores can have a chance to settle. I used to do 15 minutes because that's what I read, but I had a lot of contamination problems until I just started waiting longer. Also try to minimize the number of times you bring your hands in and out of the box.
If you're using ball jars like me, get your lid rings broken loose before you let it sit, because it's tough to break them loose in the box with your hands covered in alcohol and slippery. You wanna have everything placed right and a plan in your head of exactly what you're gonna do, do it quick and minimize the time you have the jars open.
Most people around here flame their scalpel blade between cultures, but I just use a new one so I'm not bringing my hands in and out of the box with hot blades and creating air currents. They're like $7 for a box of 100.
I'm no agar expert these are just a few things I've done to get much better results in my SAB. Also, I use the upside down box on top of moist towel routine that Notahacker420 shows here, this way any spores that fall stick to the towel and don't get stirred up again.
Good advice Ill keep the exposure to a minimum
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Adustus
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Quote:
Amanita virosa said: Although jars work fine, consider investing 7 bucks in a sleeve of sterile plastic petri plates. MUCH easier to work with, esp. in an sab. take up less room in the fridge, less awkward to sub out of and easier to see contam. Otherwise, allowing your jars to cool with the lids slightly loose (flip the lids upside down, rubber side up) will allow excess moisture to escape. Otherwise, you may have some on the surface of the agar. Consider making some slants for long term storage of your cultures.

Petris are my next investment. If you have a good source to buy a sleeve id appreciate it my friend.
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Shu
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Re: Starting my mushrooms in some agar [Re: Adustus]
#19256350 - 12/10/13 04:55 PM (10 years, 1 month ago) |
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Move very slowly in the SAB, air currents are the enemy.
Open your plates only far enough to pass in your culture and close them quickly.
Practice your transfers using empty containers and no cultures to build up muscle memory.
Transferring smaller pieces of culture to many separate dishes increases your odds of a clean transfer.
Have a few spare plates ready for second pass in case all your plates contaminate and pass the culture at the first sign of contam.
Never open a moldy plate in your SAB.
What "lid" are you speaking of?
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Adustus
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Re: Starting my mushrooms in some agar [Re: Shu]
#19264172 - 12/12/13 03:08 AM (10 years, 1 month ago) |
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Quote:
Shu said: Move very slowly in the SAB, air currents are the enemy.
Open your plates only far enough to pass in your culture and close them quickly.
Practice your transfers using empty containers and no cultures to build up muscle memory.
Transferring smaller pieces of culture to many separate dishes increases your odds of a clean transfer.
Have a few spare plates ready for second pass in case all your plates contaminate and pass the culture at the first sign of contam.
Never open a moldy plate in your SAB.
What "lid" are you speaking of?
Meant lids from my jars.
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SkysTheLimit
A curious mind


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Re: Starting my mushrooms in some agar [Re: Shu]
#19264225 - 12/12/13 03:55 AM (10 years, 1 month ago) |
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Quote:
Shu said: Move very slowly in the SAB, air currents are the enemy.
Open your plates only far enough to pass in your culture and close them quickly.
Practice your transfers using empty containers and no cultures to build up muscle memory.
Transferring smaller pieces of culture to many separate dishes increases your odds of a clean transfer.
Have a few spare plates ready for second pass in case all your plates contaminate and pass the culture at the first sign of contam.
Never open a moldy plate in your SAB.
What "lid" are you speaking of?
I think you should try spraying down the glovebox with 10% bleach or lysol or w/e after you put all your work inside. The water vapour traps dust and spores causing them to stick to surfaces. After an hour or so, you can assume most of the dust/spores in your SAB has either settled down or stuck to a surface.
I do this and I'm pretty sure most SAB users do the same, its also done in plant cloning where the specimens are just as vulnerable to contamination in mycology.
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bjanuszewski
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Re: Starting my mushrooms in some agar [Re: SkysTheLimit]
#19265141 - 12/12/13 10:24 AM (10 years, 1 month ago) |
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if your worried about contam you need to make sure you have clean tissue thats been handle very little. you can always isolate away from contam too.
my greatest success with agar comes from using 1/2 pint jars w/ tyvek over AE hole and using my colonized rye berries to inoc the agar. using tissue can be hard bc you'll usually have to isolate to get nice clean looking mycelium from my experience.
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Adustus
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Registered: 11/14/13
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Re: Starting my mushrooms in some agar [Re: Adustus]
#19270911 - 12/13/13 02:01 PM (10 years, 1 month ago) |
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Well it seems to have gone well so far. I spray like all hell through every step with 91% alcohol or peroxide. My gas exchange tape melts to my jars in the PC and I think my rack got all fucked up too.
Do i need more water in my PC? I fill it to just cover the jar lids.
Anyone have any ideas for gas exchange? I tap two holes with nails in the lid and planned to use the tape but it doesnt seem to like the PC. Will leaving the holes open allow too much moisture inside?
I just need to figure out my gas exchange now :/
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blindingleaf
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Re: Starting my mushrooms in some agar [Re: Adustus]
#19271227 - 12/13/13 03:16 PM (10 years, 1 month ago) |
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if u r using 1/2 pint glass jars u can just turn the lid disc upside down (even that is not needed though), agar barely need GE (though it may be debated here, I have found it does not) I think you should search "Doc-T's no pour Tek" because he uses 1/2 pint glass jars and if I remember correctly, no MOD for GE, just put the law disc and ring on normal with no holes. if u already have holes in the lid, go with Tyvec like bj said, but don't cook them with exposed holes, WAAAY too much moisture will get in... u don't need more water in PC, less than if u are cooking grains because the PC will only run 30-35 minutes. also, don't put too much agar in each jar, because its annoying to transfer something thicker than 1/4 inch, I'd say go for 1/8 inch, u don't need much, just a light covering of the bottom. In my AA if I put water to JUST under the rack disc it came with, thats more than enough to run 30-35 minutes. Good Luck! Agar is fun. I think Im better with agar than actually growing shrooms! its addicting, get some PP5s and ill have a full time hobby, as if growing wasn't enough right?
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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blindingleaf
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Re: Starting my mushrooms in some agar [Re: blindingleaf]
#19271235 - 12/13/13 03:17 PM (10 years, 1 month ago) |
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also, my micropore tape is fine in PC, esp if just for 30 min, is that the tape u were referring to that did not do well in PC?
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Adustus
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Re: Starting my mushrooms in some agar [Re: blindingleaf]
#19273560 - 12/14/13 06:24 AM (10 years, 1 month ago) |
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Quote:
blindingleaf said: also, my micropore tape is fine in PC, esp if just for 30 min, is that the tape u were referring to that did not do well in PC?
Yeah I got some weird cheap shit I found online. Well Im new to agar and I made way too much from what it sounds. I bet my reishi culture I traded for is fucked. Ill have to review that tek and take another shot at agar. Do you use food coloring in yours to spot contams better?
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Forrester
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Re: Starting my mushrooms in some agar [Re: Adustus]
#19273628 - 12/14/13 07:09 AM (10 years, 1 month ago) |
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If the tape isn't working out, you could always use post-office tyvek. Holds up well in the PC. Just silicon a little patch over whatever hole you've got in the lid.
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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Adustus
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Re: Starting my mushrooms in some agar [Re: Adustus]
#19273683 - 12/14/13 07:47 AM (10 years, 1 month ago) |
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Badass! I just found out about the pillow stuffing as a means of gas exchange. I have a couple tubes thatll do fine.
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Forrester
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Re: Starting my mushrooms in some agar [Re: Adustus]
#19273895 - 12/14/13 09:37 AM (10 years, 1 month ago) |
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Yeah polyfill works too. Just cover the lids with foil in the PC.
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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Amanita virosa
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Re: Starting my mushrooms in some agar [Re: Adustus]
#19273903 - 12/14/13 09:41 AM (10 years, 1 month ago) |
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Quote:
Adustus said: Well it seems to have gone well so far. I spray like all hell through every step with 91% alcohol or peroxide. My gas exchange tape melts to my jars in the PC and I think my rack got all fucked up too.
Do i need more water in my PC? I fill it to just cover the jar lids.
Anyone have any ideas for gas exchange? I tap two holes with nails in the lid and planned to use the tape but it doesnt seem to like the PC. Will leaving the holes open allow too much moisture inside?
I just need to figure out my gas exchange now :/
Too much water! Two inches of water is all you need in the pc. Just be sure to vent the air out till pure steam is coming out before putting the weight on. If you put holes in the lid and the tape melted they are toast. Micro pore tape makes a terrible filter. Honestly, you could leave out the gas exchange altogether. Just use plastic lids. They are not air tight. Or even better buy some dang petri plates. You can re sterilize the agar. It will liquefy when heated.
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t3chnobily
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Re: Starting my mushrooms in some agar [Re: Forrester]
#19273910 - 12/14/13 09:42 AM (10 years, 1 month ago) |
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pack the polyfill tight if you want it to be an effective filter. I use tyvek squares taped down with 3m micropore and they last many pc cycles.
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blindingleaf
blue collar underworld



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Re: Starting my mushrooms in some agar [Re: t3chnobily]
#19275145 - 12/14/13 03:11 PM (10 years, 1 month ago) |
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amanita...I have been curious to try not putting a ge holes in these containers..

I was under the impression that without one they would pop in the PC...do u think the pressure inside would just slowly push the air out instead, even though its a snap on? for the screw on PP5 I just loosen them a bit, no GE hole.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Amanita virosa
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Re: Starting my mushrooms in some agar [Re: blindingleaf]
#19275217 - 12/14/13 03:30 PM (10 years, 1 month ago) |
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Dunno. Have you tried nuking them? If the are not polypropylene or some comparable plastic they will melt. I suspect the lids would pop right off.
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blindingleaf
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yea they are def. pp5, not tried nuking, don't own a microwave 
the reason I have been curious about NOT modifying these snap on PP5s is because the lid is loose enough that when u spin it, there is no resistance. but it must be air "restrictive" enough that no con tams enter...I have these (the PP5 in my last post) just sitting in my dining room with no contam, I smoke (in adjacent room, with door closed but drafty perimeter), have 2 cats and a dog that are within inches of my jars and PP5s..and they remain clean, even long term, like 3-4 weeks, like when I use them for mini grain isolate testers. I don't wanna jinx myself and say the L word...but...
any thoughts?
i might as well try it anyway, just need to get more, or wait till the ones I have grow enough to transfer.
random question to anyone... after u guys take transfers from a plate, whether petri, PP5, etc, do u generally just toss that sender plate? or mark it, and put in fridge for longer storage if indeed u got results that were intriguing...? if so, does this increase contam risk?
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
Edited by blindingleaf (12/14/13 04:16 PM)
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Adustus
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Now only if my damn roommate would wake up so I can bang some holes in these jars and get PCing my grains and agar. Ive only done my reishi so far. And even though I think I did a piss poor job my first time around vs contams(leaving jar open longer than I wanted). It seems the little guys fighting the good fight. How long till I can spot contams?
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blindingleaf
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Re: Starting my mushrooms in some agar [Re: Adustus]
#19276088 - 12/14/13 07:02 PM (10 years, 1 month ago) |
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no telling when I guess. are you talking about con tams with agar? if so, then if they got on the plate while pouring or inoculating they will grow as fast or faster than the myc. if u r talking about grain jars/brf they can show at any time. i am unfamiliar with reishi myc and how it should/should not look, though I would def be interested in trying to grow it! PM me if u wanna trade something for it! I've never traded before, but I want to. I have actives in print form (just b+ and soon burma and peurto rican) and shiitaki and oyster cultures, both isolates. I don't even know how one might send cultures in the mail, but if i find out and u wanna trade, we should! I live in philly, so from here to new york is not too far! Im glad I posted on this thread, it seems like we both may be at the same level in the hobby experience wise, so it makes for good experience sharing btw us! maybe in five years we will be on Amanita virosa and forresters level giving ppl like us now good advice! we will grow up together on the shroomery haha!
ps... try this thread, there are a bunch of pages of just ppl posting pics of agar plates with edible and actives, I like looking through it occasionally when I'm bored, so maybe there will be some reishi pics on it. good luck! http://www.shroomery.org/forums/showflat.php/Cat/0/Number/8328896/an//page//vc/1
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Adustus
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Registered: 11/14/13
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Last seen: 1 year, 6 months
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Re: Starting my mushrooms in some agar [Re: blindingleaf]
#19276702 - 12/14/13 09:42 PM (10 years, 1 month ago) |
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Quote:
blindingleaf said: no telling when I guess. are you talking about con tams with agar? if so, then if they got on the plate while pouring or inoculating they will grow as fast or faster than the myc. if u r talking about grain jars/brf they can show at any time. i am unfamiliar with reishi myc and how it should/should not look, though I would def be interested in trying to grow it! PM me if u wanna trade something for it! I've never traded before, but I want to. I have actives in print form (just b+ and soon burma and peurto rican) and shiitaki and oyster cultures, both isolates. I don't even know how one might send cultures in the mail, but if i find out and u wanna trade, we should! I live in philly, so from here to new york is not too far! Im glad I posted on this thread, it seems like we both may be at the same level in the hobby experience wise, so it makes for good experience sharing btw us! maybe in five years we will be on Amanita virosa and forresters level giving ppl like us now good advice! we will grow up together on the shroomery haha!
ps... try this thread, there are a bunch of pages of just ppl posting pics of agar plates with edible and actives, I like looking through it occasionally when I'm bored, so maybe there will be some reishi pics on it. good luck! http://www.shroomery.org/forums/showflat.php/Cat/0/Number/8328896/an//page//vc/1
Once its grown id love to send you a live culture on the house man. Ive only got my first agars going. Lol. So I'm prepping all my jars now and im using polyfil for gas exchange. Should i be using a clump big enough to cover the top or just the hole? And are they reusable?
On the note of how to send live cultures one member on here traded me for 8 live cultures in one go and sent me a small envelope fulled with mini plastic vessels with pop tops. He labeled each one with a roman numeral and included a ID card for each number. Shit was boss!
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blindingleaf
blue collar underworld



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Re: Starting my mushrooms in some agar [Re: Adustus]
#19277925 - 12/15/13 06:35 AM (10 years, 1 month ago) |
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sweet! how big were the pop top tubes?
as far as poly, Ive never used it as a jar/agar filter, only used in mono tubs/ bucket grows. I know it does not cover the whole lid, just the GE hole. and probably reusable, but its so cheap that I would replace each time for peace of mind.
http://www.shroomery.org/forums/showflat.php/Number/18278624
that might be helpful for getting an image of how the poly should look with lids. are u talking about poly for grain jars? that tek also has an injection port, but u won't need it because u are doing an agar wedge inoculation correct? so just follow that tek instruction on the poly part of the lid, and then ur good to go! cover with foil during PC, and I even put a rubber band around foil below the lip of the jar so no steam hits anything.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Amanita virosa
botanist by day


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Re: Starting my mushrooms in some agar [Re: blindingleaf]
#19278334 - 12/15/13 09:22 AM (10 years, 1 month ago) |
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Quote:
blindingleaf said: yea they are def. pp5, not tried nuking, don't own a microwave 
the reason I have been curious about NOT modifying these snap on PP5s is because the lid is loose enough that when u spin it, there is no resistance. but it must be air "restrictive" enough that no con tams enter...I have these (the PP5 in my last post) just sitting in my dining room with no contam, I smoke (in adjacent room, with door closed but drafty perimeter), have 2 cats and a dog that are within inches of my jars and PP5s..and they remain clean, even long term, like 3-4 weeks, like when I use them for mini grain isolate testers. I don't wanna jinx myself and say the L word...but...
any thoughts?
i might as well try it anyway, just need to get more, or wait till the ones I have grow enough to transfer.
random question to anyone... after u guys take transfers from a plate, whether petri, PP5, etc, do u generally just toss that sender plate? or mark it, and put in fridge for longer storage if indeed u got results that were intriguing...? if so, does this increase contam risk?
I usually keep them around but I am working in front of a hood. I like to watch them for clues about contam. Since we sell cultures, I will often use a clean plate to fill orders over the course of a month. Another strategy that works well is to transfer wedges from your plate into individual micricentrifuge tubes. They come sterile. You can make thirty from one plate if ur careful. Them when you need a fresh plate just dump the wedge onto one.
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blindingleaf
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Registered: 07/19/13
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wow, never heard of those tubes! r they small? need to be kept in fridge? available on vendor sites? is that how shroomery members send live edible cultures in the mail for trading?
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Forrester
aspiring sociopath


Registered: 02/05/13
Posts: 9,351
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Re: Starting my mushrooms in some agar [Re: blindingleaf]
#19279738 - 12/15/13 02:48 PM (10 years, 1 month ago) |
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Quote:
blindingleaf said: wow, never heard of those tubes! r they small? need to be kept in fridge? available on vendor sites? is that how shroomery members send live edible cultures in the mail for trading?
They're what a lot of people use for trading, they work great for it. Yeah, they're pretty small (1.5ml, I think there's other sizes too). Outgrow has them, that's where I got mine. A lot of other places carry them too.
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blindingleaf
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Re: Starting my mushrooms in some agar [Re: Forrester]
#19280154 - 12/15/13 04:41 PM (10 years, 1 month ago) |
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awesome, gotta check them out. how is the culture preserved when in the mail? is it just a small wedge that is usually mailed with h2o or something to that effect?
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Forrester
aspiring sociopath


Registered: 02/05/13
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Re: Starting my mushrooms in some agar [Re: blindingleaf]
#19280253 - 12/15/13 05:02 PM (10 years, 1 month ago) |
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Yep just an agar wedge, dropped into the tube, no water.
-------------------- Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here. ------------------- Have some medicinal mushrooms and want to get the most out of them? Try this double extraction method.
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Adustus
Multiple Personalities




Registered: 11/14/13
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Re: Starting my mushrooms in some agar [Re: Forrester]
#19282429 - 12/16/13 04:39 AM (10 years, 1 month ago) |
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Whats cool is they help reduce contams when inoculating a jar. Just pop it open and drop it in. Minimal exposure. Love it! And I can reuse the ones I have now to send live cultures to other people too 
I got my agar jars and my grain jars fitted with the GE filter using poly. PCing 3 jars of agar now. My reishi seems to be putting up a great fight against my epic fail with my agar. No contams so far but its taking a long time to take over. Is this normal?
I forgot the food coloring now that I think of it. But itd be stupid to stop it now and put it in. I should have a decent time spotting contams with the shroomery on my side though. Maybe I should just learn to spot contams on normal agar.
Fuck all this though im off to get some plates next time I get some money.
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