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ManifoldPrime
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Help Manifold diagnose slow spawn colonisation
#26840907 - 07/23/20 03:35 PM (3 years, 6 months ago) |
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For the longest time I have struggled with slow colonisation times. I feel like this is the biggest hurdle I currently face, with half of my grain spawn taking at least 2 months to reach an iffy 99%, and half not even making it to 60% in that time.
For those of you that remember Van Hatton's "Help me create clean spawn" thread, I would like to make this similar thread, not only to log trying different methods and changing variables to increase colonisation times for myself, but for others that have similar issues to read.
I used to mess around with LC / LI back when I first started, and while my contam rates were high, when something was clean it would amaze me with how fast it would run and reach 100%. I dont do LC anymore, but I still feel like my agar wedge inoculation is wayyyy too slow. While I am going to start getting back into LC's and such as well, I want the focus of this thread to be based on inoculation with relatively large agar wedges.
Here are some pics of my current batches of spawn, with variety and inoculation date/ current colonistation time noted.
RW I dont see this variety working wonders on plates and cakes, but its still competitive and for fucks sake these were inoculated as far back as 6 weeks ago (10/6) Some are so close to being done, but have sections that are sloooooowly closing up.
 These were agonisingly behind, and were shaken yesterday.

MZ Performs well on agar, but I've yet to fruit this so it's an unknown variable at this point. Seems strong on some other small masters and such. inoculated 17/6. 5 weeks ago.
   
PESA The most consistent variety I own, usually fast and aggressive on plates and cakes. inoculated recently on 26/6. 4 weeks ago. Not too bad, but I feel like it should be over 50% at this point.


Variables that affect colonisation time, and my current status/understanding/relationship with them:
- Temperature. I keep my spawn on a shelf in a reasonably cool room. It is winter now, but I've always had this issue even during the heat of summer. Right now the temperature guage right above the spawn jars reads:

- Grain prep. The examples posted are all relatively new grains for me to prep, and I followed Ham/Bod's teks to the T, but they are following the same time scale as the WBS/Millet spawn I also have running at the same time.
- Grain hydration. I suspect I've prepped the rye correctly but the long colonisation time and FAE has them currenty (as of posting) on the dryer side.
I made small 100ml controls with these batches, perhaps I should just cut up some of those grains and check the hydration in the center?
- Shaking. I shake vigorously after noccing up, and usually at around 30%, and again at 90-100% for confirmation on recovery time. However my recovery time is usually piss poor, taking around 3-4 days to revert to original percentage complete. Should I be shaking more intervals? I dont see the point.
- Genetics. I've got different varieties on agar, and most of them are from last year. I have freshly germinated varieties on grains right now that are more or less in line with the examples shown on some pints. I would understand if genetics or slow cultures of these varieties was at fault if there was at least 1 that was sailing ahead, but every jar to be inline with other spawn as if they are all set to the same slow rate.
- Contamination, especially bacteria. I am extremely vigilant on my agar transfers, and my transfer plates/ no-pours rarely have any contamination show up. I should also note that I dont really see a shoebox contaminate, even well after second flush. Those RW oats look iffy but the rye looks squeeky clean to me.
-------------------- My Magnum Opus:Thai Variety Comparison Please correct me if I say something wrong! | Current Trade Print Availability:Some PESA and RW available for South African posters. Snail Mail only. |
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Tight Lunchbox
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Re: Help Manifold diagnose slow spawn colonisation [Re: ManifoldPrime]
#26840949 - 07/23/20 04:06 PM (3 years, 6 months ago) |
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How long do you pc your oats? Your RW look bacterial to me, but I'm not familiar with oats so I'm probably not the best judge.
Do you have any pictures of the cultures on agar?
-------------------- "it's all a joke between mom contractions and coffin fittings" The most useful tool for noobs
Edited by Tight Lunchbox (07/23/20 04:18 PM)
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natedawgnow
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Re: Help Manifold diagnose slow spawn colonisation [Re: Tight Lunchbox]
#26840974 - 07/23/20 04:22 PM (3 years, 6 months ago) |
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Some of those grains look like oats and some like rye.
In the first set of pics, oats I assume, they look bacterial for sure. Not a fan of oats for lots of reasons, but they are usually harder to get moisture right and those jars look wet and bacterial.
As for the rye pics, those grains look extremely dry. The pic I'm about to post is of wheat, but wheat and rye are very similar grains so i think it's relevant:

The grain on the left is what a fully hydrated grain should look like. The grain on the right is a dry grain. Your grains look more like the grain on the right.
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ManifoldPrime
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Re: Help Manifold diagnose slow spawn colonisation [Re: natedawgnow]
#26842597 - 07/24/20 12:21 PM (3 years, 6 months ago) |
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Quote:
How long do you pc your oats?
Those were done for at least 90 minutes at 15PSI. Usually I vent, let the pressure hit 15PSI, start the timer, and when the 90 minutes are up, slowly knock back the heat over 5 minutes, which usually keeps the pressure over 10PSI for around 10-15 minutes. I cant fit more than 7 of those specific bottles in the PC so the thermal load is low per cook. I recently started overclocking to 17 PSI. I'll kick up the time to 2 hours for the next batch.
Quote:
Your RW look bacterial to me, but I'm not familiar with oats so I'm probably not the best judge.
I'll concur but do note I did a control of each of those grains, and all of them look squeaky clean.
Quote:
Do you have any pictures of the cultures on agar?
Not of these specific cultures I dont think. I've more or less finished with those specific strains, but I may have other plates in the same lineage lying around. I'll check and document.
For the next batch of grain I do I will take pics of everything involved.
Quote:
those grains look extremely dry
I investigated the rye control jars and yes, they are probably dry. The grains were on the threshold between mushy and powdery. Its odd, I boiled and strained as per the simmer tek. I will time every step of the prep down to the second for the next run, hopefully I can get it done tomorrow. Should I also do a batch of Soak grains? I also did a batch of sorghum/millet masters and they look properly hydrated but those grains were soaked.
-------------------- My Magnum Opus:Thai Variety Comparison Please correct me if I say something wrong! | Current Trade Print Availability:Some PESA and RW available for South African posters. Snail Mail only. |
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mushboy
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Re: Help Manifold diagnose slow spawn colonisation [Re: ManifoldPrime]
#26842607 - 07/24/20 12:31 PM (3 years, 6 months ago) |
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I do not like to shake after inoculation regardless of inoculate used.
Makes it tricky to diagnose contamination sources.
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sandman420
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Re: Help Manifold diagnose slow spawn colonisation [Re: mushboy]
#26842806 - 07/24/20 02:24 PM (3 years, 6 months ago) |
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None of the RW looks like cubensis mycelium to me. Some kind of white mold or hella bacterial or sumthn. So you are doing good with the other strains but your RW is all contaminated. Probably a bad print or syringe that you started from I'd bet. Or those jar lids on those type of jhars could be whack...
Edited by sandman420 (07/24/20 02:45 PM)
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sandman420
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Re: Help Manifold diagnose slow spawn colonisation [Re: sandman420]
#26842820 - 07/24/20 02:35 PM (3 years, 6 months ago) |
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What do your jar lids look like? I just now see that those others were started 5+ weeks ago thats no bueno.
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ManifoldPrime
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Re: Help Manifold diagnose slow spawn colonisation [Re: sandman420]
#26843039 - 07/24/20 04:30 PM (3 years, 6 months ago) |
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Quote:
None of the RW looks like cubensis mycelium to me
The exact same lineages that went into those jars were more or less proved on no pours and cakes:

Its cube myc but probably bacterial.
Quote:
just now see that those others were started 5+ weeks ago thats no bueno.
Those are starting to bruise blue - probably from dehydration. I'm gonna chuck them.
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I do not like to shake after inoculation regardless of inoculate used.
Good advice. when do you shake then? I suppose a week is a good window to determine if a contamination has germinated elsewhere from the inoculation site.
Quote:
What do your jar lids look like?
I'll upload a pic if need be, but its basic stuffed tight polyfill in a hole thats made with a nail, covered with a square of micropore for neatness/prefiltering.
Tomorrow I'll do a batch of oats or rye, dealers choice, and document everything, procedure and all. I dont have any specific culture lined up, I'll pick based on my observation of which plates are ready. How big a transfer should I do? I usually make a 2-3cm2 wedge. Most of my cultures right now are on Grainwater agar, and look clean - should I only use the plates that are MEA?
-------------------- My Magnum Opus:Thai Variety Comparison Please correct me if I say something wrong! | Current Trade Print Availability:Some PESA and RW available for South African posters. Snail Mail only. |
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ManifoldPrime
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Re: Help Manifold diagnose slow spawn colonisation [Re: ManifoldPrime]
#26844972 - 07/25/20 05:32 PM (3 years, 6 months ago) |
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Spent a good deal of time today cooking both Oats and Rye, in half litre masons: 
Followed Bod's oat and Ham's rye teks to the T. Timers and everything. 2 hours at 15-17 PSI.
Oatz:
 
Rye:
 
They look a lil wet coming out the PC. I find they dry a bit after cooling.
Example of the Polyfilled+taped top lid:

Also cooked some nopours to save the cultures I transfer tomorrow. Will update tomorrow with inoculation procedure and culture pics.
-------------------- My Magnum Opus:Thai Variety Comparison Please correct me if I say something wrong! | Current Trade Print Availability:Some PESA and RW available for South African posters. Snail Mail only. |
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mushboy
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Re: Help Manifold diagnose slow spawn colonisation [Re: ManifoldPrime]
#26844993 - 07/25/20 05:52 PM (3 years, 6 months ago) |
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They might dry a bit but those grains look way to wet imo.
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Tight Lunchbox
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Re: Help Manifold diagnose slow spawn colonisation [Re: mushboy]
#26845261 - 07/25/20 10:12 PM (3 years, 6 months ago) |
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Same^
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LotKid
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Re: Help Manifold diagnose slow spawn colonisation [Re: Tight Lunchbox]
#26845563 - 07/26/20 06:18 AM (3 years, 6 months ago) |
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Yup
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ManifoldPrime
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Re: Help Manifold diagnose slow spawn colonisation [Re: LotKid]
#26846131 - 07/26/20 01:20 PM (3 years, 6 months ago) |
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-------------------- My Magnum Opus:Thai Variety Comparison Please correct me if I say something wrong! | Current Trade Print Availability:Some PESA and RW available for South African posters. Snail Mail only. |
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mushboy
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Re: Help Manifold diagnose slow spawn colonisation [Re: ManifoldPrime] 1
#26846144 - 07/26/20 01:26 PM (3 years, 6 months ago) |
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I'd let your culturs grow out more and take a few more transfers. None of those look ready to use yet.
Just my opinion.
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LotKid
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Re: Help Manifold diagnose slow spawn colonisation [Re: ManifoldPrime] 2
#26846154 - 07/26/20 01:29 PM (3 years, 6 months ago) |
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ManifoldPrime
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Re: Help Manifold diagnose slow spawn colonisation [Re: LotKid]
#26846174 - 07/26/20 01:42 PM (3 years, 6 months ago) |
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@mushboy yeah those cultures were all T2-T3. I do have other cultures on hand that are further down the line in terms of transfer count, but I recently transfered like, all of them and werent grown out enough. also nocced up two of those spawn jars with KSSS germ plates As you can see in that vid I was doing transfers of those donors at the same time, as well as just general transfers of everything else I want on spawn. Next round of spawn next weekend, gonna probably do more oat jars and noc them up with RW and more Thais that are T3-T4. I wanna dial in the moisture. I'm just not sure why I cant seem to get oats dry enough?
@lotkid Amon Tobin, Autechre, Aphex, Lorn, Jon Hopkins, Cern, IDM and experimental electronic suits cult work so fuckin well 
I also found my smaller Milo and Pearl Millet masters have recovered from a shake 2-3 days ago; how are these looking?
 
I made those because I felt that I need to get over my fear of G2G. I intend to G2G the best looking of those, around 1 master to 2 donor jars, maybe quarts of oats.
Edited by ManifoldPrime (07/26/20 01:45 PM)
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mushboy
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Re: Help Manifold diagnose slow spawn colonisation [Re: ManifoldPrime]
#26846220 - 07/26/20 02:26 PM (3 years, 6 months ago) |
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I dried my oats to what i thought was way to dry and that's when they were perfect. Like, stupid dry. Dry dry.
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sandman420
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Re: Help Manifold diagnose slow spawn colonisation [Re: mushboy]
#26846478 - 07/26/20 05:18 PM (3 years, 6 months ago) |
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why's that millet so grey?
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ManifoldPrime
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Re: Help Manifold diagnose slow spawn colonisation [Re: sandman420]
#26847070 - 07/27/20 02:23 AM (3 years, 6 months ago) |
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Quote:
I dried my oats to what i thought was way to dry and that's when they were perfect. Like, stupid dry. Dry dry.

Quote:
why's that millet so grey?
Because its not actually millet, its pearl millet, a grain often grown in africa.
-------------------- My Magnum Opus:Thai Variety Comparison Please correct me if I say something wrong! | Current Trade Print Availability:Some PESA and RW available for South African posters. Snail Mail only. |
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Drboomer
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Re: Help Manifold diagnose slow spawn colonisation [Re: mushboy]
#26855467 - 07/31/20 12:31 PM (3 years, 6 months ago) |
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Quote:
mushboy said: I dried my oats to what i thought was way to dry and that's when they were perfect. Like, stupid dry. Dry dry.
This, I hated oats until recently. I under hydrate with a 20ish minute boil them,let them dry with a fan on them until they look dry dry then PC. The little bits of excess moisture makes them nice and plump
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