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OfflineDabrit
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Agar no signs of germination after 3 weeks toss?
    #23913116 - 12/10/16 01:26 PM (7 years, 1 month ago)

So I inoculated 4 agar plates with ms solution( B+) 3 weeks ago now and literally there are no signs of growth at all be it bacterial or mycelium. Used Pasteywhyte's agar tek/recipe.

At this point should I just accept that likely my spore syringes are dead and toss em? ( Over a year old now).

Tried direct inoculating WBS with no signs of growth either after a month and tried two different B+ syringes which arrived at the same time.

Perhaps my storage of the syringes wasn't sound?? Kept them in a black plastic bag in the fridge caps were put back on the needles.

Still waiting on my new PE & GT syringes to arrive.


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Psychedelics = True gateway to spiritual enlightenment and detachment from ones meat costume :wink:




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OfflineHuskies
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23913160 - 12/10/16 01:40 PM (7 years, 1 month ago)

Is this your first time with agar? My first time I was dumb and I put the plates with my grain jars, they were in for 90 minutes. It cooked them too much I think, and neither spores or bacteria ever took hold on it.


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I call them Huskies cause you tell them to go "Mush! Mush""


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OfflineWeavieWonder
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23913165 - 12/10/16 01:42 PM (7 years, 1 month ago)

Could be bunk.  It happens.  Start fresh with your new syringes.


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InvisiblemushboyMDiscord
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Re: Agar no signs of germination after 3 weeks toss? [Re: WeavieWonder]
    #23913171 - 12/10/16 01:44 PM (7 years, 1 month ago)

1 yr old spores in the fridge is kinda old. prints last awhile. not sure about
spores inside a dirty fridge :x


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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: Huskies]
    #23913176 - 12/10/16 01:45 PM (7 years, 1 month ago)

Quote:

Huskies said:
Is this your first time with agar? My first time I was dumb and I put the plates with my grain jars, they were in for 90 minutes. It cooked them too much I think, and neither spores or bacteria ever took hold on it.




Yeah first time trying out agar. I guess the fact that it's not showing any signs of bacteria is maybe at least showing me I'm sterilizing the agar/plates correctly. I'm using MS tape as a filter no wrap being used on the outside of the plates as per using glad mini rounds. Storing at room temperature out in the open on a table in my room.


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Psychedelics = True gateway to spiritual enlightenment and detachment from ones meat costume :wink:




Edited by Dabrit (12/10/16 03:39 PM)


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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: mushboy]
    #23913179 - 12/10/16 01:46 PM (7 years, 1 month ago)

Quote:

WeavieWonder said:
Could be bunk.  It happens.  Start fresh with your new syringes.



Quote:

mushboy said:
1 yr old spores in the fridge is kinda old. prints last awhile. not sure about
spores inside a dirty fridge :x





Yeah kinda what I was figured. Hopefully the new syringes arrive any day now but maybe not given it's christmas time and they are shipping over the border and I just went with the regular shipping option.


--------------------
Psychedelics = True gateway to spiritual enlightenment and detachment from ones meat costume :wink:




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OfflineHuskies
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23913184 - 12/10/16 01:48 PM (7 years, 1 month ago)

How long did you have them in the pressure cooker for? My first batch looked too much like the sugars caramelized or something. I had also cooked it into a paste on the stovetop, instead of pouring it in quite liquid.


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I call them Huskies cause you tell them to go "Mush! Mush""


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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: Huskies]
    #23913263 - 12/10/16 02:12 PM (7 years, 1 month ago)

Quote:

Huskies said:
How long did you have them in the pressure cooker for? My first batch looked too much like the sugars caramelized or something. I had also cooked it into a paste on the stovetop, instead of pouring it in quite liquid.




40 minutes


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OfflinePanarchist
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23913444 - 12/10/16 03:17 PM (7 years, 1 month ago)

if you didn't see any growth whatsoever it's possible those syringes were filled with nothing but water.


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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: Panarchist]
    #23913450 - 12/10/16 03:20 PM (7 years, 1 month ago)

Quote:

Panarchist said:
if you didn't see any growth whatsoever it's possible those syringes were filled with nothing but water.




Nah I could see the spores in the syringe. And they came from a reputable site sponsor.


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Psychedelics = True gateway to spiritual enlightenment and detachment from ones meat costume :wink:




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OfflinePinPornProducer
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23913474 - 12/10/16 03:28 PM (7 years, 1 month ago)

Try leaving the syringe out in room temp for a couple weeks and try again


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Offlineenlightenment
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23913477 - 12/10/16 03:29 PM (7 years, 1 month ago)

Strange... It should work... What is your agar recipe? How do you inoculate the agar? What is the temperature?


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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: PinPornProducer]
    #23913497 - 12/10/16 03:33 PM (7 years, 1 month ago)

Quote:

PinPornProducer said:
Try leaving the syringe out in room temp for a couple weeks and try again




Thanks I'll try that


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Psychedelics = True gateway to spiritual enlightenment and detachment from ones meat costume :wink:




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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: enlightenment]
    #23913506 - 12/10/16 03:34 PM (7 years, 1 month ago)

Quote:

enlightenment said:
Strange... It should work... What is your agar recipe? How do you inoculate the agar? What is the temperature?




I'm using PasteyWhyte's agar recipe. inoculated the agar plates inside of a SAB.

Temp in room 22 degrees Celcius


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Psychedelics = True gateway to spiritual enlightenment and detachment from ones meat costume :wink:




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Offlineenlightenment
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23913528 - 12/10/16 03:40 PM (7 years, 1 month ago)

Everything sounds right so far... Maybe the syringe was exposed to very high/low temperatures for some time. :shrug:
Wait for the other syringe and try to message the vendor to ask for a replacement.


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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: enlightenment]
    #23913548 - 12/10/16 03:46 PM (7 years, 1 month ago)

Quote:

enlightenment said:
Everything sounds right so far... Maybe the syringe was exposed to very high/low temperatures for some time. :shrug:
Wait for the other syringe and try to message the vendor to ask for a replacement.




who knows they've been in my fridge since last November except for when they were taken out to knock up some jars. Last active May of this year.

Funnily enough the bonus syringe they came with of Amazonian spores contaminated all 8 of my PF Tek jars as well with cob web mold.  Although who knows what happened there could have been myself messing up somehow with my sterile technique, as it was only my 2nd time doing a grow at the time.


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OfflineKenetic
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23913566 - 12/10/16 03:52 PM (7 years, 1 month ago)

Id reuse the agar too.  I had about 15 'duds' as I call them due to a shitty technique (put some spore solution on a piece of foil and rubbed my loop around in it blindly) and I simply reused the plates for something else.

Of course I go through a decent amount of agar and dont keep plates after I tranfer from them. 


--------------------
Todo Cambia
   

               
                                                :cookiemonster::elmo:



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Offlineenlightenment
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23913569 - 12/10/16 03:53 PM (7 years, 1 month ago)

OK. If they are one year old it is too late to contact the vendor.
I used 1.5 year old syringes from the fridge without problems..
Sterile technique looks OK. You should see contaminations on agar if it would have been bad.

It looks like the only possibility is to wait for your syringes and start over. Good luck.


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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: Kenetic]
    #23913589 - 12/10/16 03:58 PM (7 years, 1 month ago)

Quote:

kenetic said:
Id reuse the agar too.  I had about 15 'duds' as I call them due to a shitty technique (put some spore solution on a piece of foil and rubbed my loop around in it blindly) and I simply reused the plates for something else.

Of course I go through a decent amount of agar and dont keep plates after I tranfer from them. 




Yeah I guess it really doesn't matter too much about sterilization on your first inoculation as you are basically planning on contaminates being present and cleaning it up anyway.

Honestly I think I'm just gonna do a fresh batch of agar and use the actual petri dishes that arrived the other day. It's really not that much harder to pour the plates and use regular petri dishes as oppose to the Glad mini rounds. Seen as tt is kinda hard to see into the glad mini round containers especially if there is any condensation present and the lids are blue and I being a genius used blue food coloring in my agar mix lol.


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Psychedelics = True gateway to spiritual enlightenment and detachment from ones meat costume :wink:




Edited by Dabrit (12/10/16 04:03 PM)


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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: enlightenment]
    #23913623 - 12/10/16 04:06 PM (7 years, 1 month ago)

Quote:

enlightenment said:
OK. If they are one year old it is too late to contact the vendor.
I used 1.5 year old syringes from the fridge without problems..
Sterile technique looks OK. You should see contaminations on agar if it would have been bad.

It looks like the only possibility is to wait for your syringes and start over. Good luck.




Yuup can't wait for them to arrive. I have a bunch of WBS sitting waiting for when the time comes. Then once that's all gone I think I'm gonna start working with rye seed and see if I can find a local farmer who can give me a good deal on it. Seems pretty pricey ordering it online up here above the border.


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Psychedelics = True gateway to spiritual enlightenment and detachment from ones meat costume :wink:




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OfflineKenetic
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23913656 - 12/10/16 04:13 PM (7 years, 1 month ago)

Quote:

Dabrit said:

Seen as tt is kinda hard to see into the glad mini round containers especially if there is any condensation present and the lids are blue and I being a genius used blue food coloring in my agar mix lol.




:rofl:

About reusing them I do it because I'm lazy and that shit's not cheap.  If they're clean, they're clean.  I use them before they dry out anyway.

As far as pour agar being easy, it's actually easier than most people think.  I can use agar way better than I can actually grow mushrooms lol

Btw syringes suck on agar, i much prefer using prints


--------------------
Todo Cambia
   

               
                                                :cookiemonster::elmo:



DMT said: Everyone know's me, they just don't know it yet


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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: Kenetic]
    #23913675 - 12/10/16 04:17 PM (7 years, 1 month ago)

Quote:

kenetic said:
Quote:

Dabrit said:

Seen as tt is kinda hard to see into the glad mini round containers especially if there is any condensation present and the lids are blue and I being a genius used blue food coloring in my agar mix lol.




:rofl:

About reusing them I do it because I'm lazy and that shit's not cheap.  If they're clean, they're clean.  I use them before they dry out anyway.

As far as pour agar being easy, it's actually easier than most people think.  I can use agar way better than I can actually grow mushrooms lol




Hey man you don't need to justify shit to a noob like me lol.


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Offlineenlightenment
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23913773 - 12/10/16 04:39 PM (7 years, 1 month ago)

Quote:

Btw syringes suck on agar, i much prefer using prints




True if you have fresh prints. But you should not generalize this theory.

I had a 5 yr old Na Muang print that did not germinate on agar by swiping it with a sterile swab on several dishes. After making a spore syringe to rehydrate the spores for ~1 week 5 of 5 dishes showed growth.

Spore syringes work well on agar if you do it right. I recommend o use an inoculation loop. Flame the inoculation loop, let it cool by sticking it into the receiving agar dish. Push out one drop of spore solution on the inoculation loop and do this:



(You can skip step 3-5 if you do not have a flow hood because it takes some time to make all of these steps. That adds a higher contam risk)


Edited by enlightenment (12/10/16 04:44 PM)


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OfflineKenetic
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Re: Agar no signs of germination after 3 weeks toss? [Re: enlightenment]
    #23913865 - 12/10/16 04:56 PM (7 years, 1 month ago)

Shouldnt you sterilize between each swipe?


--------------------
Todo Cambia
   

               
                                                :cookiemonster::elmo:



DMT said: Everyone know's me, they just don't know it yet


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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: enlightenment]
    #23913882 - 12/10/16 05:00 PM (7 years, 1 month ago)

Quote:

enlightenment said:
Quote:

Btw syringes suck on agar, i much prefer using prints




True if you have fresh prints. But you should not generalize this theory.

I had a 5 yr old Na Muang print that did not germinate on agar by swiping it with a sterile swab on several dishes. After making a spore syringe to rehydrate the spores for ~1 week 5 of 5 dishes showed growth.

Spore syringes work well on agar if you do it right. I recommend o use an inoculation loop. Flame the inoculation loop, let it cool by sticking it into the receiving agar dish. Push out one drop of spore solution on the inoculation loop and do this:



(You can skip step 3-5 if you do not have a flow hood because it takes some time to make all of these steps. That adds a higher contam risk)




Nice little demonstration you got there. Will keep that in mind :smile:


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InvisibleComebackKid
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23914168 - 12/10/16 06:24 PM (7 years, 1 month ago)

If you're using agar agar powder it's likely that you are using too much agar.
Pasty tek calls for 2.5 grams
Try 2 or 1.8 grams for softer agar. Will help with germination.

Also try making brf paste and using that to germinate in your plates.

Probably just stubborn spores.
I got a ksss print that was only weeks old. I was using pasty recipe and went through ~16 plates with no action and finally germinated on 1/5 brf paste plates.


--------------------
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Look around you... Everything you see exists inside the mind.
Consciousness, the awareness that is experiencing this mind,
is peering in from outside the universe.

Our individual experiences are all part of the universe's experience of itself


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OfflineDabrit
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Re: Agar no signs of germination after 3 weeks toss? [Re: ComebackKid]
    #23914198 - 12/10/16 06:32 PM (7 years, 1 month ago)

Quote:

ComebackKid said:
If you're using agar agar powder it's likely that you are using too much agar.
Pasty tek calls for 2.5 grams
Try 2 or 1.8 grams for softer agar. Will help with germination.

Also try making brf paste and using that to germinate in your plates.

Probably just stubborn spores.
I got a ksss print that was only weeks old. I was using pasty recipe and went through ~16 plates with no action and finally germinated on 1/5 brf paste plates.




Wouldn't BRF paste make it harder to see bacterial infections as opposed to your standard agar?


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InvisibleComebackKid
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Re: Agar no signs of germination after 3 weeks toss? [Re: Dabrit]
    #23917029 - 12/11/16 05:07 PM (7 years, 1 month ago)

Yes
That's why you transfer the myc to agar after to clean it up :thumbup:


--------------------
:amanita2: Substrate surface conditions / Monotub prep and care :sporedrop:

Look around you... Everything you see exists inside the mind.
Consciousness, the awareness that is experiencing this mind,
is peering in from outside the universe.

Our individual experiences are all part of the universe's experience of itself


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