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Moower
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Time to start - log? 1
#26637231 - 04/29/20 04:58 PM (3 years, 8 months ago) |
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After some previous experience with cakes over a year ago, with so so results, I decided to prep myself much more before next attempt at growing. Future journals I will place properly at my private profile, but for now I hope for some helpful insights from time to time as it's my first bulk grow 
First things first.
Agar plates: For those I decided to run with BOD's Agar Tek. 1.9% Dark Malt Extract | 1.9% agar agar | 96.2% H20. Poured a total of 38 plates before I run out of agar. During whole process I was keeping plates sheltered in the sleeve, and decided to secure the leftovers by knotting the sleeve for later. I know it might be contam factor for later projects, but lets roll a dice for now. 28 plates went to freezer, 10 were introduced to spores using inoculation loop within SAB.
I have run into some condensation problems, probably because of pouring too hot . Lesson learned for future.
         
After 2 days no sign of growth, neither good nor bad, so i'll roll with this as a rather good sign and wait patiently. I was getting used to new medium so I experimented with force needed (caused some bruises for example on PR2), but from what I've been reading here it shouldn't cause problems.
For further processes I'm aiming for: -A+ Baq tek for preparing grain, as well as Manure free substrat for Pan Cyan; -Blenderless Liquid Inoculation straight to bags with patches; -BOD's unBODified Monotub Tek with little modification to the type of boxes available localy
Edited by Moower (04/30/20 12:31 AM)
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ModularMind
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Re: Time to start - log? [Re: Moower]
#26637277 - 04/29/20 05:19 PM (3 years, 8 months ago) |
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Any idea when the print was taken?
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Moower
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Circa 2 months ago - to the paper, then each individually stored in zip lock baggie. They are from a private vendor with a lot of positive reviews so I'm in a green when it comes to their viability.
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ModularMind
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Re: Time to start - log? [Re: Moower]
#26637953 - 04/30/20 12:17 AM (3 years, 8 months ago) |
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Yeah. No worries. Just curious. Looking forward to updates.
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c10h12n2o
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Hang tight
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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Moower
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Re: Time to start - log? [Re: c10h12n2o]
#26641398 - 05/01/20 01:54 PM (3 years, 8 months ago) |
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As of now there is still no visible change.
C10h12n20 - thanks for stepping by, gif in your signature was the method I've used to distribute spores 
I've made some more reading during last 2 days and came into possible problem. After I PC my grain, the bags probably will be air-tight. I'll need them to be opened tho to transfer myc using blenderless technique... I guess SAB won't be sterile enough for that? Or will be? Flow hood unfortunately is not an option right now.
Can anyone point me in the direction of the TEK showing how to properly do it? At start I thought that I'll just open them in SAB, pour liquid inside and use zip-lock to close them again, but this thread made me realize how foolish that was of me.
Is it even possible? Should I revert my plan to using jars?
Edited by Moower (05/01/20 01:56 PM)
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c10h12n2o
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Re: Time to start - log? [Re: Moower] 1
#26641960 - 05/01/20 06:36 PM (3 years, 8 months ago) |
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Use a syringe and needle
Wipe w alcohol, inject, tape or hot glue the hole
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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psillypsally
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Re: Time to start - log? [Re: c10h12n2o]
#26642151 - 05/01/20 08:29 PM (3 years, 8 months ago) |
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Quote:
c10h12n2o said:

lmao
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Moower
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Re: Time to start - log? [Re: c10h12n2o]
#26642429 - 05/01/20 11:42 PM (3 years, 8 months ago) |
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Quote:
c10h12n2o said: Use a syringe and needle
Wipe w alcohol, inject, tape or hot glue the hole
No need for some air for colonization?
Wouldn't parts of agar clog the needle? Should I go for meat injection one (3mm) instead of regular 1.6mm?
Separate syringe for every agar plate, or cleaning by pumping 70% ISO few times between inoculations should be enough?
Edited by Moower (05/02/20 12:00 AM)
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c10h12n2o
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Re: Time to start - log? [Re: Moower] 1
#26642623 - 05/02/20 01:58 AM (3 years, 8 months ago) |
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You can make LI or LC first. I use a standard syringe and 18g to 20g needle
I love LCs , lots of people prefer LI tho.
Before you inject the bag, hang it for a few minutes to an hour, it will suck air in through the filter and have enough air for colonization and easy inoculation
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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Moower
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Re: Time to start - log? [Re: c10h12n2o]
#26647297 - 05/04/20 01:00 AM (3 years, 8 months ago) |
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A week since inoculation and we can see some change. I'm happy to observe and learn that different species truly show different behavior when it comes to colonizing.
First to turn white were TN2 dish - on Saturdays morning, TN1 followed thru till Saturdays evening. For AA+ today's night was a breaking point, and it turned from semi-transparent white to having few patches of white. I believe that PR will follow today. And both MK still show no sign of growth, neither good of bad.
 
Alongside update I came again with couple of questions:
-I've got a word from my vendor that PP bags are 65 microns thick, and the filter patch retains particles up to 0.1 micron both in air as well as in water and technological steam. Should I seal them before pressure cooking or after? I'm going to use zip-locks for that.
-Biggest syringes I can get in bulk for relatively small price are 20ml. When it comes to inoculating with LI i've read that it should be done with 20-25ml / kg of grain, so obviously I'll have to poke each bag 3x, given I'm going to fill them with 3kg of grain. Obviously different syringe for different species will be a must, but I wonder If my sterile technique will be sufficient for using the same syringe for 3 fills or should I open a new one for each fill?
Open syringe in SAB, flame sterilize the needle and wipe syringe with 70% ISO, open jar with LI, fill syringe, close jar, poke hole in the receiving bag, squirt inside, tape the hole, wipe needle, syringe and hand with ISO and flame the needle, repeat procedure peeling off the same piece of tape for each fill.
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ModularMind
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Re: Time to start - log? [Re: Moower]
#26647309 - 05/04/20 01:10 AM (3 years, 8 months ago) |
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Have you considered applying silicone to the bags as a self healing injection port?
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Moower
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I haven't. Doesn't silicone need at least some grip on surface to stay put?
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c10h12n2o
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Re: Time to start - log? [Re: Moower]
#26647343 - 05/04/20 01:40 AM (3 years, 8 months ago) |
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Opening the jar over and over is gonna mess you up. Ideally use a jar with a ship for LC /LI
Tape works fine. Ive had silicone come off bags, seems to work for other people tho... i prefer tape
Quick tip. Wipe bag injection site with iso, then use sharpie to draw small circle , inject in circle so the hole doesnt get lost.
.1 micron is tiny, i dunno if presealing is gonna work w that. Usually works fine w .2, .5 and .4 tho. Make sure you let the heat go down as slowly as possible if you try it
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  C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide "Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing." "Convictions are more dangerous enemies of truth than lies" ― Friedrich Nietzsche
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Moower
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Re: Time to start - log? [Re: c10h12n2o]
#26650810 - 05/05/20 01:37 PM (3 years, 8 months ago) |
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I'll ordered some rubber gaskets to fit jar leads as inoculation ports and secure them with RTV.
Will do my pest to use zip-ties after they are out of PC. I hope no contams get inside.
Some myc have grown faster, some slower.
I'm starting to thing about first series of transfers. I'd prefer to do those during the weekend. Both because of time availability and because 3 plates are behind, and 3 still shows no sign of growth (2 McKennaii's, I wonder what up with them???). So here is the question if I can wait so long till Saturday, or would it be wiser to do it on two different occassions. Especially with those AA+ and Temar Negara plates as they have gained speed.

Last 3 pictures are almost certainly myc, as they look similar to previous 4 in earlier stages of development.
Edited by Moower (05/05/20 01:42 PM)
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Moower
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Re: Time to start - log? [Re: Moower]
#26668503 - 05/13/20 11:47 AM (3 years, 8 months ago) |
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So after all I took transfers both on 6th and 10th of May.
Those are the results of transfers a weeek ago:
And those from 3 days ago.

All of them seems to be recovering nicely, although I was a little bit concerned about almost transparend mycelium on AA+ and Puerto Rico plates. On the other hand TN 2T1 plate looks super promising and I really can't wait to fruit this one.
They started to whiten yesterday. This leads me to follow'up transfers again that I'll do either tomorrow or during the weekend. The plan is to to transfer white dot from AA+ 1T1 9 o'clock, PR 1T1 10:30 o'clock and TN 2T1 5 o'clock.
Q1: Any advice regarding switching the place from which to transfer is highly welcome!
One of McKennai plates finally showed some movement, but so slow that I can't even take a good picture of it. Myc looks semi transparend / very light white.
Q2: May it be caused by agar composition? Like stated previously its ~2% Agar & ~2% DME
Today I also prepared some injection ports on lids thru which I'm planning to suck the LI to syringe for inoculation in the futue. I've experimented with rubber plug orientation to determine which one will last longer.<I've probably overused sylicone >
During next week both new pressure cooker as well as grains should arrive So i'll start playing with those and post further updates.
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Moower
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Re: Time to start - log? [Re: Moower]
#26678983 - 05/18/20 01:10 PM (3 years, 8 months ago) |
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Today my Pressure Cooker arrived. Unfortunately the seller is a moron and doesn't know what he is actually selling.

Long story short - valve weight is rated @ 0.5 bar. After doing some measuring it turns out that even this rating is overrated.
So i did some calculations. The hole threw wich the steam escapes is 6mm in diameter. Simple math = 28.27mm^2 surface. The weight is 118g on its own. This gives us circa 4,2bar. Not so good huh? So fast trip to fishing store and I got myself some additional weights for under a buck. After quick assembly my new DIY weight sits at 283g.
If my math is correct this should provide pressure of 1 Bar / 14.5 PSI when it's lightly wobbling. Fortunately 2nd security valve is rated @ 1.1 Bar so there should be no risk of explosion (as there is 3rd also), and just small margin for correct pressure.
I decided to prepare my rye today. Started by putting water to boil and then turning the heat off. Added the rye to boiling water and closed the lead for 2 hours. During this time I mixed the rye twice.After this time I've poured the rye thru colander and layered on the clean towel. Turned the fan on let let it cool and dry for 3 hours mixing occasionally. From 3000g dry, I've achieved 5389g wet product. This translated to roughly 45% water content. For next batch i'll aim for 2-2.5 hour to try to keep water content at 50%.

After that all the grain was separated into 6 bags, each of which was locked with reusable zip-tie.

Tomorrow should be the day of running it in pressure cooker.
What is Your advice regarding the time?
My plan is to stack 2 steam strainers (to allow more water inside) and then run it 3-4 hours. What You say?
P.S. I forgot about additional transfers I did yesterday. If they turn clean I plan to store them for further use. What is proper storing technique? Wait for them to colonize fully and put into freezer / fridge?
Those are three plates transferred on 06.05. As long as TN goes I'm sure it's healthy. But AA+ and PR one looks totally different - more cottony. Do they look clean to You? Asking because those are the one I'm planning to inoculate tomorrow.
Edited by Moower (05/18/20 03:14 PM)
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Moower
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Re: Time to start - log? [Re: Moower]
#26681101 - 05/19/20 12:59 PM (3 years, 8 months ago) |
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So far it looks like my "improvement" worked out It took me forever to heat up PC on my electric plate. Probably 30min of airing before just steam was escaping. I've added the weight and cooked for 3h, occasionally adjusting temperature. 2 times 2nd security valve started to release steam - thus I'm positive that pressure was around 15 PSI.
I was worried that bags would pop because of preseal - no such thing happened. They were really steamy and compressed after whole process with couple of grains popped.
I've inflated them by hanging for couple of minutes.
On the second run I've pressure cooked jars for LI.
After 3h of cooling, I've mounted my SAB and began proces of creating LI - shaked the hell of them. Agar haven't dissolved completely, but strains of myc were visible to the eye in the liquid.
Using needles I've inoculated bags and taped them using micropore tape.

Right now they sit in the closet with open doors @ 23.6C. Fingers crosed. Now the wait begin.
Edited by Moower (05/19/20 02:30 PM)
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Moower
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Re: Time to start - log? [Re: Moower]
#26708709 - 05/31/20 08:40 AM (3 years, 7 months ago) |
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12 days into and most bags were at 20-30% colonization. I've decided to shake them. Well, more like 'knead' them. This resulted in some shroomy smell being released into air thru filter. All seems good. AA+ seems to be the fastest colonizer of the rye.
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Moower
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Re: Time to start - log? [Re: Moower]
#26750595 - 06/17/20 02:47 AM (3 years, 7 months ago) |
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Almost month has passed from grain inoculation. For the last 5 days, bags seemed to reach their limits. All 6 of them looked more or less like the one below. You can notice a small patch of uncolonized grain on the bottom left corner, but leading-edge moved circa 1mm during those 5 days, and mycelium was nicely packed when I decided to crunch bags into pieces.

Yesterday evening I've prepared coir. 500ml of water per 100g of dried coir. 2.5 brick. Turned out it was a little bit to much water, so for next run, I'll aim at 450ml / 100g, as I had to squeeze some water for proper field capacity.
Each lid is reversed to introduce fruiting conditions straight away. This created gaps around handles and latches:
 
And now the wait begins:

Somewhen this week I'll hang 6500k led strips on top of each shelf with 12/12 timer.
As You can see, boxes are placed in the wardrobe. Could I close the does for proper fea or at least circle it 12h open / 12h closed (day / night)?
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