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Invisiblepoopy mcpooperson
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agar isolate log. (PICS)
    #19013539 - 10/22/13 12:34 PM (10 years, 3 months ago)

well that was a real pain in the ass that looked like it was going to be really quick, simple, and easy. i used parafilm, 4 section petri dishes, 10 different syringes of 10 different varieties from 2 different vendors, MEA, they are sitting in a closet at 70-72 degrees, and i used lots of love...sweet love. sweet fucking love.

i think i over did it though. i used two 4 section dishes for each variety and one 4 section dish with drops from all of them. that one was for shits and giggles and really inappropriate since im trying to isolate.

photos to come.


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Edited by poopy mcpooperson (10/28/13 10:43 AM)


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. [Re: poopy mcpooperson]
    #19013676 - 10/22/13 01:03 PM (10 years, 3 months ago)

It'll be worth it in the end when you have a fat canopy


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Invisiblepoopy mcpooperson
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agar isolate log. (PICS) [Re: abltsandwich]
    #19042214 - 10/27/13 07:11 PM (10 years, 3 months ago)

ok, well since i havent found an agar start to finish log i might as well post mine. i have a few questions about my petri dishes for anyone who has agar experience. this is my first time and i think its going ok. i think the liquidy white on top of almost of these is just spore solution that didnt get absorbed into the agar and is gradually drying. the ones with white liquid either do not smell or smell like fresh mushrooms. let me know if you have any experience with MS solution doing this to agar. i am using homemade MEA agar. the ones with yellow got tossed out a minute ago. i think i just need to go easy with the spore solution next time. i used 1-2 drops for each section, needle-less just the syringe. i think next time i should use the syringe with a needle and only one drop. it might dry up quicker on the agar. i also need to not tip them so the spore will come from just one spot.

all are almost 5 days into the process, have been kept in a room at 72-74 degrees, are wrapped in parafilm, have 12 hour on/off 6300K light, and were made with love.

they are all laid out so they can have even amounts of environmental factors. i dont know if stacking them would help them or hurt them.

ARGENTINA MS SYRINGE:

these got thrown away because they smelled like an old band-aid. they all came from the same syringe which makes me think the syringe is contaminated. it smells 100% completely bacterial and i think im going to mix in antibiotics into a bit of agar before pouring into my petri dishes next time just for kicks to see if it can combat the bacteria.




COLORADO MS SYRINGE:

i think they are doing ok. i think the white is soon to be mycelium and i can see individual spots where i think there is going to be growth. these have the white liquidy solution on top.





ALA MS SYRINGE:

these have the white liquidy look to the tops of them and dont smell through the parafilm.




CREEPER MS SYRINGE:

these seem to be doing the best. they smell like fresh mushrooms and they are seem to be the fastest growing.




CAMBODIAN MS SYRINGE:

the same white liquidy top as a lot of the other ones. the dishes dont smell.



TEXAS YELLOW CAP MS SYRINGE:

i think these are doing second best. they seem to be starting myc formation.




PAN CYAN MS SYRINGE:

these seem like they are yellowing but dont smell.



GOLDEN TEACHER MS SYRINGE:

these seem to have that liquidy MS syringe fluid still on top.





this i just a funny photo of a guy who was dressed as the joker for halloween and got arrested for a DUI in Maine this year.


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love and laughter  :laugh:


Edited by poopy mcpooperson (10/27/13 10:20 PM)


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OfflineSynKyd
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19042350 - 10/27/13 07:39 PM (10 years, 3 months ago)

watching this, thanks for logging it!  :cheers:


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Invisiblepoopy mcpooperson
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: SynKyd]
    #19042712 - 10/27/13 08:53 PM (10 years, 3 months ago)

thanks, i just hope this is what its supposed to look like and that i am headed in the right direction. im trying to start up this project/hobby as properly and as fool proof possible. there is tons of info about agar prep and isolating growth after myc has colonized on the agar but not much info in-between those two points.


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19046441 - 10/28/13 02:50 PM (10 years, 3 months ago)

I think everyone gets carried a little carried away with the spores on their first agar attempt. Just a dab will do you as I am sure you are seeing.

I have never used a spore syringe to inoculate a dish. I have always used a print. When using a print I flame sterilize the inoculation loop (or scalpal) until red hot. Then I touch it into the receiving dish to cool it. This also causes some agar to get on the end of your loop. Having agar on the end of your loop allows you to touch your print and get just a few spores to stick. You then simply touch them to the center of the dish. If you can see the spores you have probably put too many. You want just a few spores right in the middle.

I'm not really sure how you would achieve this with a syringe since even a drop will contain 100s of spores in most syringes.

What you are going to have to do is let them grow out a little more and pick your best looking growth and get out your scalpal and make some transfers. Just take your scalpal and cut a little wedge of your best looking growth and set it right in the middle of the dish. I usually place it face down.

When your second transfer starts growing out you'll probably start seeing some sectoring. If I were you I would take the best looking sector from your second transfer and use it to inoculate your grain master.

It won't be a monoculture but what I would call a limited culture. I don't like using a monoculture from spores. It is too easy to get a shitty one. I would rather use a limited culture and grow it out and use the best looking cluster to clone for good genetics

You should also take plenty of spore prints for next time. I think you'll like using a print to inoculate your dishes more than a syringe

I hope this helps. Just holler if you need anything else :smile:


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Invisiblepoopy mcpooperson
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: Stromrider]
    #19046539 - 10/28/13 03:13 PM (10 years, 3 months ago)

ok, i have some prints that i just got in and will definitely be trying them out next, per your advice, but i have so many damn MS syringes that i dont know what do with besides trying to place them on agar. i went a little overboard when i started this project and probably own 16 MS syringes and 8 prints. I dont know what the hell i was thinking. i just got all excited and internet click happy, i guess.

in conjunction with the MS syringes:
1) i was thinking of making my agar with a little less than the recommended water amount to see if the MS syringe solution would become absorbed into the drier agar.

2) i was also thinking of making the MEA recipe verbatim again and taking my sterile scalpel and digging out a divot of agar for the MS syringe solution to nestle into real nice like in the center.

i will use the prints that i have to make some agar plates like you mentioned. i just want to be able to use the MS syringes on agar and isolate so i know for certain im not wasting my time with putting them directly on grain. if i had inoculated a bunch of jars with the ARGENTINA syringe i would had had a whole batch of smelly dirty band-aids laying around. barf.

thanks for the advice on the print to agar.


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OfflineStromriderM
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19046568 - 10/28/13 03:20 PM (10 years, 3 months ago)

Making a little divot to put a drop of spore solution in is not a bad idea. Use a red hot scalpal to do it though.

Also remember after you've been a member here for 90 days you'll have access to the marketplace and those vendor syringes will be great barter to obtain different Cultivation supplies and exotic prints :thumbup:


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: Stromrider]
    #19046575 - 10/28/13 03:21 PM (10 years, 3 months ago)

Hey Poopy, got your message to check this out.

When I read you were starting your agar attempts with spore solution, the first thought that ran thru my head is "time to see a whole bunch of bacteria-infected plates." I see I was right.

Although they often work out just fine on grains, I've never gotten spore solutions to work out on agar any better than yours except ones I made myself from my sterile spore prints.
For that reason I suggest going straight from spore solution to grains.  Here is a tek I came up with to do this and yet still have petri-style growth, with many other advantages too. They only take 2-3 drops each! Syringes last essentially forever (as in 3 drops can turn into unlimited prints)


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19046691 - 10/28/13 03:38 PM (10 years, 3 months ago)

I agree with Stromrider

Here's a link to how I make agar.

Next time, use the needle so you can flame sterilize it. Open and attach in your SAB/flowhood.

When using agar, less is more. You only need a drop.

It's hard to tell from your pics, but your agar either looks like it doesn't cover the bottom of the plate and/or it's not mixed well.

Yeah, liquid runs when you tilt the dish. That's one reason to use only a drop or start using prints.

In the RR video, He actually wipes the spores around in a tiny circle with his loop. This also makes sure the spores are seated. It's how I do it.

I'm about to use a syringe for agar for the first time. I've only used prints and myc in the past. I may use my loop the same way as for prints.

But just like with prints, MS to agar often requires a transfer or so to isolate away from the contamination. That's the main reason I got in to agar.


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Invisiblepoopy mcpooperson
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: Violet]
    #19046699 - 10/28/13 03:40 PM (10 years, 3 months ago)

so you think all of the sections are bad? they dont seem yellow to me and they dont stink bad expect for that first batch. will the white turn to yellow or is that just excess water on the surface from the syringe like when you get wrinkled skin after swimming for too long?

i tried to click on that link but it took me nowhere.


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: Violet]
    #19046713 - 10/28/13 03:42 PM (10 years, 3 months ago)

Quote:

Violet said:
Hey Poopy, got your message to check this out.

When I read you were starting your agar attempts with spore solution, the first thought that ran thru my head is "time to see a whole bunch of bacteria-infected plates." I see I was right.

Although they often work out just fine on grains, I've never gotten spore solutions to work out on agar any better than yours except ones I made myself from my sterile spore prints.
For that reason I suggest going straight from spore solution to grains.  Here is a tek I came up with to do this and yet still have petri-style growth, with many other advantages too. They only take 2-3 drops each! Syringes last essentially forever (as in 3 drops can turn into unlimited prints)




I can't believe took so long for someone to mention the fact that pretty much all of those plates are contaminated.  (I kind of skimmed some of the posts, so I apologize if I missed something.)  OP - Sorry to say, but those all look pretty rough.  I agree with Violet; use spore solution to inoculate some grain jars, and from there it's pretty easy to pull out a couple grains and put them on a dish.  I've also had much better success with that than liquid spore solution.  Maybe it has something to do with the fact the dish is a fairly small environment, so that little bit of extra moisture does a lot of harm; I have noticed that any dishes that end up with condensation seem to have more of a tendency to contaminate than those that don't. 

Anyway, I'd toss those and start fresh.  You'll love it once you get some that grow well; it's so cool watching mycellium grow on a flat surface.


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Invisiblepoopy mcpooperson
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: SpitballJedi]
    #19046741 - 10/28/13 03:46 PM (10 years, 3 months ago)

the wheels are rolling along in my head now. you got me thinking, spit.

what if i used, lets say pressure cooked mason jar lids, and put it in my SAB. i could put a drop or two from my syringe onto the cooled mason jar lid and then transfer to the agar with my inoculation loop. this might be the answer. im going to do a batch with all three ways i have posted, in addition to the print TEK, and see how they do.

at first, all i wanted wanted was a tub of mushrooms to stick my face in and graze on like a cow feeds on grass but it has become much more than that at this point. this might consume me if im not careful.


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19046757 - 10/28/13 03:50 PM (10 years, 3 months ago)

Just the lid? How would you keep it sterile?

If you want to go along that route, look up "no pour agar" tek.

Before you switch horses, give your method another try. Some things just take a little practice.


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Edited by SpitballJedi (10/28/13 03:51 PM)


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Invisiblepoopy mcpooperson
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: illuminati]
    #19046773 - 10/28/13 03:54 PM (10 years, 3 months ago)

shit. that was my worry. they dont stink though. do you even think that the creeper and TYC are contaminated? they seem to have myc growth.

i dont think im gonna throw them out just yet as i might have a good day to day pictorial of what not to do if they are contaminated.

if i did the grain agar transfer would you suggest i do them in pint jars instead of quart jars to colonize? would i have to wait for the whole jar to colonize or just enough for me to pluck a grain easily to move to agar?

i am going to use franks grain to agar TEK and dunk the grain because it seems to have a much more rapid growth.


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19046801 - 10/28/13 04:00 PM (10 years, 3 months ago)

Even contaminated plates can still be viable if you can get a wedge cut away from the contam


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: Stromrider]
    #19046856 - 10/28/13 04:08 PM (10 years, 3 months ago)

:whathesaid:


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Invisiblepoopy mcpooperson
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: SpitballJedi]
    #19046907 - 10/28/13 04:19 PM (10 years, 3 months ago)

spit-
i could put the lids in a PC approved plastic container with a cover and then wait for the PC to cool and place the container with the lids in my SAB. i could then add the spore solution to each of the separate lids from each of the different syringes one after another taking each lid out of the SAB before moving on to my next lid. i would then use a heated loop to grab the most minimal amount possible and add to my agar. this would crate WAY less excess fluid on top of the agar and i wouldn't have to screw with my agar recipe.

maybe ill just try it with one first and see if its even worth a shit. i can always do what strom said and just trade them in the marketplace after my probationary period is over.

am i thinking about this too hard?

adderall will be the death of me.


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19046930 - 10/28/13 04:24 PM (10 years, 3 months ago)

Toss everything but the Creeper.  Sorry, but those other plates are beyond saving.


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: grainbrain]
    #19046968 - 10/28/13 04:29 PM (10 years, 3 months ago)

^Exactly

Personally I'd toss the Creeper too, and start with all of them on grains.

Since there's a good chance the syringes are actually contaminated (every one of your plates!) I'd only inoc 1 thing from each, and I suggest those be small...


--------------------
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19046997 - 10/28/13 04:35 PM (10 years, 3 months ago)

....and then put the lids back in the plastic container? Sound s like a mess.

Just use one syringe and do 1 or 2 plates. Transfer good myc to a new plate and toss the rest.

In some ways, I'm a sheep on a bandwagon. I do things that I know work for most people most of the time.

I'm willing to help you through a process I'm familiar with, but at this point, you're kind of on your own.

Take pics, post results, good luck.


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19047016 - 10/28/13 04:36 PM (10 years, 3 months ago)

Quote:

poopy mcpooperson said:
shit. that was my worry. they dont stink though. do you even think that the creeper and TYC are contaminated? they seem to have myc growth.

i dont think im gonna throw them out just yet as i might have a good day to day pictorial of what not to do if they are contaminated.

if i did the grain agar transfer would you suggest i do them in pint jars instead of quart jars to colonize? would i have to wait for the whole jar to colonize or just enough for me to pluck a grain easily to move to agar?

i am going to use franks grain to agar TEK and dunk the grain because it seems to have a much more rapid growth.





You definitely could do them in pint jars if you want, but personally, I'd rather be left with a healthy quart of spawn in addition to getting a few plates inoculated.  Either way, wait for it to fully colonize before opening it. 

Frank definitely knows a hell of a lot more about this hobby than I do, so I'd think you'd be just fine following one of his teks.


--------------------
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: illuminati]
    #19066753 - 10/31/13 10:37 PM (10 years, 2 months ago)

so i think i can isolate these dishes for transfer, right?

these were the only dishes that i kept.  can anybody let me know which areas to cut out? i have watched the agar isolate section from my RR videos and i have viewed a few old threads to try to figure it out and im still confused. can someone who has experience let me know if i am on the right track or not?

also, if i do isolate and the rest of the wedge is not contaminated would it be ok to drop it in a jar of grain or WBS?

DISH 1: im thinking bottom right at 8 o'clock


DISH 2: im thinking bottom right dish at 1 and 3 o'clock



DISH 3: i dont even know if this is mushroom myc. maybe bottom left 8 o'clock.


DISH 4: i was thinking 2 o'clock top right and 4 o'clock bottom right


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19069932 - 11/01/13 01:29 PM (10 years, 2 months ago)

any thoughts on which sections to isolate with these dishes?


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19074286 - 11/02/13 09:41 AM (10 years, 2 months ago)

can anyone tell me if where im guessing to sector for an isolate is correct?

also, if i do isolate and the rest of the wedge is not contaminated would it be ok to drop it in a jar of grain or WBS?


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: grainbrain]
    #19074674 - 11/02/13 11:23 AM (10 years, 2 months ago)

Quote:

grainbrain said:
Toss everything but the Creeper.  Sorry, but those other plates are beyond saving.




:foshizzle:

plates 1 & 2 look good to me, i'd wait and see whats happening with 3. 4 looks like its turning green in the bottom left. there's plenty of sectoring to take transfers from on plates 1 & 2, you've got the right idea. as for your 2nd question about inoculating a grain jar with leftover growth i wouldn't see why it would hurt, but if it was me i would just wait. before too much longer you're going to be inoculating a bunch of jars testing out separate isolations to see which one is best.

you're on the right track dude just hang in there! i wish i had started out working with agar instead of waiting so long and having so many crappy grows from chance genetics. i've been at it for a little over a year and am just picking up the agar work. before you know it you'll have a beautiful canopy of brown hats and so many mushrooms.. they'll be falling out of your ass.


Edited by PirateSwazey (11/02/13 11:52 AM)


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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: PirateSwazey]
    #19074785 - 11/02/13 11:53 AM (10 years, 2 months ago)

thanks so much.

i am trying to isolate and want to get clean, strong, and wonderfully producing cultures to work with.

lets say i use dish 2, which you said would be ok to isolate from. there are 4 separate agar sections on the petri. i think the bottom right of that dish looks the best to isolate from. i was just wondering if i could take the left over myc in that section, after i take out my pieces for isolation, and drop the entire leftover section it in some grain and if it would perform.

while im isolating i would like to see if i could get some type of growth, as small as it may be, so i can essentially practice my method of spawning/fruiting before i get my isolates where is want them.


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InvisiblePirateSwazey
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: poopy mcpooperson]
    #19075049 - 11/02/13 01:02 PM (10 years, 2 months ago)

Quote:

poopy mcpooperson said:
lets say i use dish 2, which you said would be ok to isolate from. there are 4 separate agar sections on the petri. i think the bottom right of that dish looks the best to isolate from. i was just wondering if i could take the left over myc in that section, after i take out my pieces for isolation, and drop the entire leftover section it in some grain and if it would perform.




i would pull from dish 1 and 2. theres a small section on dish 3 in the bottom left section that looks promising if you'll give it a few more days to grow out. both sides on the bottom of dish 1 have a few good sectors to transfer from. dish 2 i can see 3 good places on the bottom right section.

just remember that once you have each of these sectors isolated they will all be different substrains, and will colonize, fruit, and preform differently. some of them may preform really well and others may be total crap that you don't even want to see all the way through colonization.

the reason i am just saying to wait until you've got isolated genetics to inoculate a jar is that you're going to have so many different substrains to work with, and its only going to be another week or 2 down the road. with that time frame, if you were to inoculate a jar today, you'll be looking at 1-3 weeks of colonization time, then a g2g transfer to make up enough jars to create a monotub, which is going to take another week or 2 to colonize.

i suppose you could noc up the quart and spawn it into a little shoe box bin with 1 or 2 quarts of substrate and put that into a SGFC when its time to fruit it. if i was in your situation and dying to see some fruit bodies that is what i would probably do.

:goodluck:


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Invisiblepoopy mcpooperson
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: PirateSwazey]
    #19075075 - 11/02/13 01:08 PM (10 years, 2 months ago)

thank you very much. i thought those might some viable sections to take. i appreciate your advice.

since i first posted those photos dish 3 has grown out and could possible be the best isolate out of all of them.

have a good day.


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Offlineshaneh79
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Registered: 09/02/13
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: Violet]
    #19228238 - 12/04/13 07:00 PM (10 years, 1 month ago)

Hey Violet I am currently trying my first time isolating with agar and although its easy usiong a loop from a spore print, I'm forced to start with a syringe also. So what tek do you use to inoc grains petri dish style?  I'd appreciate any ideas..love your grows btw :smile:

Shane


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OfflinePocketRevolution
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Re: agar, ohh how i think im going to love to hate you. agar isolate log. (PICS) [Re: shaneh79]
    #19229140 - 12/04/13 09:59 PM (10 years, 1 month ago)

I like the look of dish 1 the best.  I would take samples from the bottom two sections, and transfer them to new agar plates.  Perhaps you could transfer 4 samples from each compartment to the 4 compartments of  new dishes.  Then observe the results.  The stuff along the bottom looks pretty good.

When I first looked at the pictures I thought the condensation on the lids of those plates was yeast or bacteria on the agar itself.  Good thing some more attentive members spotted the few good dishes.  You could still get some decent samples out of that.


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