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FeedYourMind
indiGlo



Registered: 03/17/14
Posts: 1,155
Loc: That One Place
Last seen: 1 day, 19 hours
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Learning to be patient...
#21412953 - 03/15/15 07:34 PM (9 years, 2 months ago) |
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So I have some B+ and Argentina in quart jars. 5 for each. This is not my first rodeo but I have never experienced growth this slow. In fact, I have 4 out of 10 jars with no growth at all. And yes, this was MS syringe from a trusted sponsor I've utilized many times before with a high degree of success. But still - no growth. Not even contams. Because of the altitiude I'm required to pc longer which I did, 2 hrs.
I had soaked my wbs 24 hours prior to pc, and then simmered until the grains were plump and tasteless. Laid out after draining and then loaded into jars. Tyvek filter with micropore tape over innoc hole, (4) 1/16 holes for GE. This has been my go to method with great success as I also innoc in a SAB per the tek. It's never failed me... Until now.
What should I do from here? Continue to be patient? I'm 2 weeks in from inoculation and this is what I have to show on my fastest jar:

And a quarter turn... This picture represents about how far my other jars have grown, not much.

And then my jars with nothing:

I'm going to make a new order with a new trusted vendor I've seen receive high praise here. I already have plenty of jars and wbs to innoc. I know I need to do some agar and I have some brf substitute going (6 jars per) but no sign of life yet in any of them.
Has anyone run into this problem before? Either way, I'm learning to let nature do its thing and not be the pushy prize winner I am.
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blindingleaf
blue collar underworld


Registered: 07/19/13
Posts: 22,008
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did u shake the jar after u inoculated with syringe?
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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FeedYourMind
indiGlo



Registered: 03/17/14
Posts: 1,155
Loc: That One Place
Last seen: 1 day, 19 hours
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Quote:
blindingleaf said: did u shake the jar after u inoculated with syringe?
I did not. It's perplexing, I know.
What do you think the deal is here?
Edited by FeedYourMind (03/15/15 08:18 PM)
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blindingleaf
blue collar underworld


Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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its been a really long time since i injected grains with a syringe, but, i would wait it out. they look clean otherwise, like u mentioned. i would shake when it doubles it current size and see what happens.
try using pints as masters next time. whether its a spore syringe, agar wedge, LI, or LC, its faster to colonize, so u can g2g earlier, and spawn quicker.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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FeedYourMind
indiGlo



Registered: 03/17/14
Posts: 1,155
Loc: That One Place
Last seen: 1 day, 19 hours
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Quote:
blindingleaf said: its been a really long time since i injected grains with a syringe, but, i would wait it out. they look clean otherwise, like u mentioned. i would shake when it doubles it current size and see what happens.
try using pints as masters next time. whether its a spore syringe, agar wedge, LI, or LC, its faster to colonize, so u can g2g earlier, and spawn quicker.
Yeah, I was starting anew and had to go that route unfortunately. I will def do the pint idea! I have some here so I'll make it a point to do a few of them as well as I will be gathering supplies tomorrow to do a more reliable and proven agar tek.
And yes! I'll be sure to shake. I figured it would be too small to do now, so you answered my next question. Thank you!
One last question though - what do you propose I do with the jars without growth?
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blindingleaf
blue collar underworld


Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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the jars w/o growth, not sure. i've seen people say to resterilize if there is no growth, but also no contam. i never had that issue, so maybe wait for someone else to post. i usually am too paranoid to do that, but in ur case, it might be reasonable, since there isn't anything growing anyway, good or bad.
i know u mentioned it, buy i would try really hard to get some agar going, be it no pour plates, or standard pour plates. it will help eliminate a lot of variables, with both MS or clones.
happy cultivating brother!
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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mushpunx
Fungus Punk



Registered: 04/20/14
Posts: 13,394
Last seen: 4 days, 17 hours
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Quote:
blindingleaf said: its been a really long time since i injected grains with a syringe, but, i would wait it out. they look clean otherwise, like u mentioned. i would shake when it doubles it current size and see what happens.
try using pints as masters next time. whether its a spore syringe, agar wedge, LI, or LC, its faster to colonize, so u can g2g earlier, and spawn quicker.
Yea I actually didnt consider that, using a pint to inoculate a spawn bag Ive been just pouring 1 quart into two bags. But I have a hundred pint jars upstairs from when I had a smaller PC.... awesome! Right on!
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 Amateur Mycologists United AMU Q&A
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FeedYourMind
indiGlo



Registered: 03/17/14
Posts: 1,155
Loc: That One Place
Last seen: 1 day, 19 hours
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Quote:
blindingleaf said: the jars w/o growth, not sure. i've seen people say to resterilize if there is no growth, but also no contam. i never had that issue, so maybe wait for someone else to post. i usually am too paranoid to do that, but in ur case, it might be reasonable, since there isn't anything growing anyway, good or bad.
i know u mentioned it, buy i would try really hard to get some agar going, be it no pour plates, or standard pour plates. it will help eliminate a lot of variables, with both MS or clones.
happy cultivating brother!
You too my friend! +5 for the help. 4 jars are resterilizing in pc for two hours and money deposited; about to order some syringes from a different vendor as well as plates and all my agar material.
Let's do this motha truckas.
Edited by FeedYourMind (03/15/15 11:17 PM)
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TheEaglesGift
The Nagual


Registered: 04/10/11
Posts: 10,554
Loc: Ixtlan, Mexico
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Let them be and see how they turn out.
I'd order some new syringes and get a new project going ASAP. It helps to have multiple projects going on at once so that when one doesn't go well, the others will keep you cheered up and engaged. I hate down time and disappointments.
Learn agar. 
I really wish someone had drilled into me to start agar when I was starting out, would've saved me a lot of time, wasted materials, and energy.
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Boland



Registered: 01/12/15
Posts: 16
Loc: United Kingdom
Last seen: 9 years, 1 month
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I had a similar situation waiting an age for growth (I had one jar which was growing much faster also)
Turned out the temperature in my house was to low, winter time in England with a dodgy boiler is pretty cold
I moved them to a mates house and they have started growing a lot quicker Still slow compared to previous grows, but all have growth and a after a month a couple are around 50/60%
I moved them after having a month of no growth on a couple and very little growth on the others
Might or might not be the issue
P.S. if temperature is cold the top of a fridge is a pretty decent place to keep them
-------------------- I want to die peacefully in my sleep like my grandfather, not screaming in terror like his passengers
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Achillita
Back to the basics



Registered: 05/26/14
Posts: 4,565
Last seen: 3 years, 3 months
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Re: Learning to be patient... [Re: Boland]
#21414828 - 03/16/15 07:48 AM (9 years, 2 months ago) |
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Some spore syringes don't come out right, even from trusted vendors. The first few times I started growing, I got 3 syringes and none of them worked. I think it might have been old spores or something weird. I eventually got a random one and it worked out and have been slowly growing my spore bank
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FeedYourMind
indiGlo



Registered: 03/17/14
Posts: 1,155
Loc: That One Place
Last seen: 1 day, 19 hours
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Re: Learning to be patient... [Re: Achillita]
#21415912 - 03/16/15 01:11 PM (9 years, 2 months ago) |
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I've learned agar and used it successfully. However, because of transportation and my access to limited supplies, I wasn't able to use MLE, agar-agar, and Petri dishes... Those have just been ordered.
I'm happy to report that being patient has its perks. I woke up this morning and all 12 of my brf substitute jars have some nice growth on them. Ready to innoc with wedges and make some isolates! 
Quote:
Boland said: I had a similar situation waiting an age for growth (I had one jar which was growing much faster also)
Turned out the temperature in my house was to low, winter time in England with a dodgy boiler is pretty cold
I moved them to a mates house and they have started growing a lot quicker Still slow compared to previous grows, but all have growth and a after a month a couple are around 50/60%
I moved them after having a month of no growth on a couple and very little growth on the others
Might or might not be the issue
P.S. if temperature is cold the top of a fridge is a pretty decent place to keep them
I believe this was part of the issue because I stuck a small, efficient space heater in a half opened closet and allowed the temps to get to 78/79F then figured out how to maintain it. My jars are loving it and growing a bit faster it seems. Thanks for the input!
Quote:
Achillita said: Some spore syringes don't come out right, even from trusted vendors. The first few times I started growing, I got 3 syringes and none of them worked. I think it might have been old spores or something weird. I eventually got a random one and it worked out and have been slowly growing my spore bank
So much truth to this. I know my syringe was made fresh as I was in contact with the vendor - and he's not one to lie. I think it could have been a combination of all of the above as well as a user error on my part.
Let me ask you kind people this: Do you flame and wipe with alchohol? I have been, but I hear and feel this gaseous sound once I inoculate into the jars. I wonder if I'm not letting it cool down enough and perhaps damaging the spores? Just a theory.
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mushpunx
Fungus Punk



Registered: 04/20/14
Posts: 13,394
Last seen: 4 days, 17 hours
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Youre talking about a needle? You can wipe with alchohol (I dont) as long as you do it *before* you flame sterilize.
Wiping afterwards will make it unsterile
--------------------
 Amateur Mycologists United AMU Q&A
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TheEaglesGift
The Nagual


Registered: 04/10/11
Posts: 10,554
Loc: Ixtlan, Mexico
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Re: Learning to be patient... [Re: mushpunx]
#21416555 - 03/16/15 03:45 PM (9 years, 2 months ago) |
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Quote:
mushpunx said: Youre talking about a needle? You can wipe with alchohol (I dont) as long as you do it *before* you flame sterilize.
Wiping afterwards will make it unsterile
This.
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FeedYourMind
indiGlo



Registered: 03/17/14
Posts: 1,155
Loc: That One Place
Last seen: 1 day, 19 hours
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Re: Learning to be patient... [Re: mushpunx]
#21418772 - 03/17/15 12:18 AM (9 years, 2 months ago) |
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Quote:
mushpunx said: Youre talking about a needle? You can wipe with alchohol (I dont) as long as you do it *before* you flame sterilize.
Wiping afterwards will make it unsterile
I've been wiping with alcohol prior to innoc and right after flaming - thinking I was adding a precautionary measure. Looks like I really was a bit backwards. Thank you!
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TheEaglesGift
The Nagual


Registered: 04/10/11
Posts: 10,554
Loc: Ixtlan, Mexico
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Alcohol=sanitary Flame=sterile
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FeedYourMind
indiGlo



Registered: 03/17/14
Posts: 1,155
Loc: That One Place
Last seen: 1 day, 19 hours
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Quote:
TheEaglesGift said: Alcohol=sanitary Flame=sterile
Dually noted. Thank you!
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FeedYourMind
indiGlo



Registered: 03/17/14
Posts: 1,155
Loc: That One Place
Last seen: 1 day, 19 hours
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And for those who were wondering..
I figured out what I did wrong - did not cook the grains long enough. I loaded some half pint jars with WBS to act as my master jars and after comparing the jars, there is a noticeable difference.
These most recent jars grains are soft and plump so I transferred some agar wedges into it and it's taking off. So if you have no contams and hardly any growth, this might be your issue to.

See the difference?
Cook ze grains!
Edited by FeedYourMind (03/17/15 09:07 PM)
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Algo
Backstrap Fever


Registered: 12/15/14
Posts: 3,857
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To be honest i bought some spores that were bunk one time from a sponser on this site. But all the others i got from them were awesome.
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FeedYourMind
indiGlo



Registered: 03/17/14
Posts: 1,155
Loc: That One Place
Last seen: 1 day, 19 hours
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Re: Learning to be patient... [Re: Algo]
#21424342 - 03/17/15 09:10 PM (9 years, 2 months ago) |
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It happens. I think it's happened twice to me now, but I believe my methods are suspect as well Time to clean up my act!
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