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InvisibleDrboomer
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Re: Josex' Poke: A different way of doing LC. [Re: Drboomer]
    #27311074 - 05/16/21 01:32 PM (5 months, 3 days ago)



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Re: Josex' Poke: A different way of doing LC. [Re: Drboomer]
    #27311364 - 05/16/21 05:29 PM (5 months, 3 days ago)

I've used something like those yea, they can work. I'd make a silicone gasket so you can screw down the nut real tight.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

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InvisibleDrboomer
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27311593 - 05/16/21 09:15 PM (5 months, 2 days ago)

Awesome man thank you


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Re: Josex' Poke: A different way of doing LC. [Re: Drboomer]
    #27316394 - 05/20/21 03:50 PM (4 months, 30 days ago)

Josex, are you familiar with Alien's Holy Grail? In that tek PC time is 1 hour - for agar, yes, but agar is like LC - and true, your tek's LC jars are not placed within another container, yet, I can pack a lot of 1/2 pints in a 23 quart Presto which to my uneducated mind would be similar in that the center jars would be surrounded/incased by jars kinda like in Alien's tek.

Would loading my Presto with 11 or 22 jars necessitate the need to increase your tek's PCg time?

If so, to what time frame?

I originally posted this question in another thread, don't know why I didn't go straight to the one who would know, although it took me awhile to coherently frame my question :foreheadslap:


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Re: Josex' Poke: A different way of doing LC. [Re: Threads2theSoul]
    #27316420 - 05/20/21 04:06 PM (4 months, 30 days ago)

I PC LME Agar and LME broths for 25 minutes at 15 psi.
If that was insufficient I should know by now after years doing the same.
Depending on altitude and media used you might have to increase times.

You can go one hour with LME if going overkill gives you peace of mind, though.
I like overkill myself but not with LME broths, because if I PC it longer than 25 minutes the liquid starts getting increasingly darker and I prefer a clear broth. I also like to take the jars out as soon as the gauge hits zero, for the same reason, the longer they stay in the PC, the darker the broth gets.

I like one hour for grain water broths, tho.

Quote:

Would loading my Presto with 11 or 22 jars necessitate the need to increase your tek's PCg time?




Nah you'll be fine, just make sure you vent well for at least 10 minutes and you'll be good, no need to increase times. Broths and agar sterilize  really easily.

You tipically only need to increase times when the internal mass of the individual vessels is bigger, but that kinda only applies to grain and denser substrates, ie. jars vs bags, where a bigger mass would require longer PC times.

Never did holy grailness but I really don't think putting the plates inside a bigger vessel would necessitate longer times.


--------------------
LC's are easy
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Clean yo shit
LAGM '21

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Edited by Josex (05/20/21 04:40 PM)


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OfflineInthepit
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27316544 - 05/20/21 05:05 PM (4 months, 30 days ago)

Hey Josex,
Quote:

I like one hour for grain water broths, tho.



Is that Oat broth from preparing oats? The broth used in agar recipies?

I've only been keeping the broth in fridge and then adding to the agar prep.


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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27316551 - 05/20/21 05:09 PM (4 months, 30 days ago)

Thanks Josex for your quick reply! I live on the coast, so pretty much just above sea level.

I was stressing about this yesterday and ran the cycle for 33 mins... why 33? Hell if I know, well, yeah I do know - for agar times per BOD's tek it's a tad over halfway between 25 and 45 mins.

AND I left them in the PC overnight, just took them out now. I had doubled your recipe and used 3.03 g LME (I doubled your PO of 1 and your likeing/using 2 and didn't want to remove that .03 to make it an even 3) and 1,000 ml distilled water. They're still clear! Although some sediment can be seen floating around on the bottom. That's normal, right? Or did I not mix up the broth well enough?

Wish me luck! This will be my first time making LC using what myc I have left from my first attempts with agar started from a liquid culture given me. At the time I was too new to know I could have used that culture to make others (I am TOTALLY new to anything shrooms). But... through that culture I was able to get it to agar and successfully use what was left making 5 rye masters to make 37 rye jars and those to make up six Pasty EZ tubs. I did take a couple of clones from that harvest.

Now... to repeat the process without loosing what's left of what I started with...


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Re: Josex' Poke: A different way of doing LC. [Re: Inthepit]
    #27316562 - 05/20/21 05:14 PM (4 months, 30 days ago)

Yea any type of grain broth. IME, one hour is  overkill with grain water (45 minutes is enough) but like I said, I like overkill if I don't break anything in the process.

@Threads2theSoul, I think I was a little vague when I said I don't think putting plates in a bigger container will require longer times.

If said vessel is filtered or isn't air tight (it shouldn't be if you are going to put plates inside), the pressure and temps inside said vessel will equalize pretty quickly.
What I mean is that the temp inside the vessel will be equal to the temp in the PC, so that's why I don't think you'd need longer times. However, overkill won't be a bad thing for LME agar so you can go longer than 25 minutes.

It's only with LME broths that I wouldn't go longer than 25 minutes because I like them clear and with no sediment.

Yes sediment is normal, especially if you left those jars in the PC overnight.

Good luck man :cheers:


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

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OfflineInthepit
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27316794 - 05/20/21 08:13 PM (4 months, 29 days ago)

Ok thanks Joesex,
If I understand you correctly, then I will PC the left over Oat broth
in a media bottle for 45+ minutes. Maybe my simply refrigerated broth is a vector?
This is the first I've heard of doing this.


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Re: The Biopsy Method: A different way of doing LC. [Re: Inthepit]
    #27318465 - 05/22/21 10:07 AM (4 months, 28 days ago)

Thank you for your write much wisdom in this thread


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Re: The Biopsy Method: A different way of doing LC. [Re: Forgottenone]
    #27318802 - 05/22/21 03:15 PM (4 months, 28 days ago)

Josex, I'm using 1/2 pint jars with unmodified grey Ball lids tightened down (no insert added to make a leakproof seal). I haven't yet done the poke and got to thinking...

How freaked out do I need to be to keep the liquid from hitting the top of the jar when I swirl it? If some gets up there and leaks to or touches the threads am I contaminating the LC? I mean, does the LC touching the threads give contams a pathway in?

Should I toss what I made, purchase inserts, and start again?

Should I have, or should I now, wrap the jars in parafilm or something else?

Or should I have faith, leave as is, take a deep breath, relax, and see what happens?


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Re: The Biopsy Method: A different way of doing LC. [Re: Threads2theSoul]
    #27318828 - 05/22/21 03:52 PM (4 months, 28 days ago)

I don't know, I've never done unmodified lids for LC.
I guess you could shake the shit out it? Although I don't think an LC ever needs to be shaken the shit out of. Swirl normally everyday and you won't have clumps so bad that would need a shake to break.

Have to say though that I'm of the strong opinion (almost Hitlerian :dancinghitler:)  that an LC lid must be filtered, not because the culture will need the GE, but for other reasons.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27318842 - 05/22/21 04:01 PM (4 months, 28 days ago)

Btw what do you mean by inserts?


--------------------
LC's are easy
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Clean yo shit
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27318965 - 05/22/21 06:24 PM (4 months, 27 days ago)

I read somewhere you can purchase like silcone rubbery rings to insert into the lid?


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Re: The Biopsy Method: A different way of doing LC. [Re: Threads2theSoul]
    #27319040 - 05/22/21 07:44 PM (4 months, 27 days ago)

I don't recommend using unmodified lids for LC but if you're going to do it anyways I think your safest bet would be a quality threaded lid, not a regular ball lid.

I think many people shy away from adding a filter to their LC lids because they think they need to silicone a watman filter to it, but this is not the case.

You can use SFD's too and there are plenty of hydrophobic filter materials on the market that you can use.
Even polyfil works great if you know how to jam it nice and tight through the whole (has a bit of a learning curve, believe it or not).

LC doesn't need to be shaken. Swirling works great and if you need something to help you break the clumps you can throw something in the jar like marbles, broken glass, blender balls, you name it...

If you swirl normally the filter doesn't need to get wet with nutrient broth, BUT I've had filters that got wet by accident and I didn't have a single issue, not even with polyfil.
For peace of mind you can choose hydrophobic filter material but you can make anything work as long as it's not some sad micropore tape or something like that.

The reason I wouldn't use unmodified lids is because, in order to PC it, you either:

  • Shut the the lid tight, hence creating a vacuum during the cycle, so when you open the lid the jar is going to suck in air (this may not be much of a problem in front of a flowhood, but it is a big risk in a SAB).
  • Leave the lid cracked during the cycle and tighten the lids immediately after taking the jars out of the PC when they're still hot. Dirty air is going to be sucked in but the really hot liquid might take care of them nasties. The problem is that if you tighten the lids when they're still hot, we still have the same problem, a vacuum is created due to the hot liquid inside.
  • Leave the lids cracked during the cycle and remove the jars the next day when they're cold.
    Again, the jars are going to be sucking in air while they are in the PC and if you don't own a sterilizer capable of holding a vacuum chances are (small but still there) that nasties can get in the jars.



The risks above are greatly minimized by making use of threaded lids, sterilizers and flowhoods. But for peace of mind and reliability, I'd always filter the lids for LC.
Also, fuck unscrewing a bulky plastic lid to inoculate the broth. Not my jam, no.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

PHENO HUNTERS


Edited by Josex (05/22/21 08:22 PM)


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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27319062 - 05/22/21 08:09 PM (4 months, 27 days ago)

Forgot to add that if you make LC's ranging from 150ml to 300ml (wise), you'll have a lot of room to swirl if you use quart jars for your LC's.

Many people (especially inexperienced people) make huge LC's of 500+ ml.
This is a bad idea because two 250ml LC (500ml total) will be ready in less than half the time it takes a single 500ml LC, but you'll still have 500ml of LC when all is said and done.

Also, a 500ml LC is not practical in most people's scenarios, you can inoculate the world with that. You could noc 250 spawn jars with a 500ml LC if you wanted, not many people need that much spawn.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

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Edited by Josex (05/22/21 08:15 PM)


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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27319112 - 05/22/21 08:43 PM (4 months, 27 days ago)

That was me, making quarts of lc from syringe like :freewilly:


Using an unmodded widemouth i had a vacuum so strong the lid bent like 45 degrees when I pried it open.


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Re: The Biopsy Method: A different way of doing LC. [Re: A.k.a]
    #27319130 - 05/22/21 09:06 PM (4 months, 27 days ago)

:flexin:

Yeah lol breaking the vacuum can be hard sometimes, especially in a SAB wearing gloves with iso.
The tip of the scalpel helps, but it's better to never create a vacuum in the first place.

Dem vacuums suck, like literally... :uhoh:


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

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Re: The Biopsy Method: A different way of doing LC. [Re: A.k.a]
    #27319156 - 05/22/21 09:28 PM (4 months, 27 days ago)

I appreciate your time Josex and hope my questions don't end up frustrating you!!! I wasn't aware you could purchase a better quality threaded lid than what comes with the mason jars and/or the others purchased separately like the (socalled) leakproof grey Ball lid. I'll look around and see what I can find.

I remember reading 'contaminated air cannot travel through obscure pathways in all circumstances' and found the post. It was in regard to agar, would it be the same for LC? Here's the link: https://www.shroomery.org/forums/showflat.php/Number/26368517#26368517. Now that I remember (my brain just doesn't compute like it used to), I'm sure that's what prompted me to go with the unmodified lids and why I was asking if liquid is introduced to the threads when swirling, would that give a pathway for the nasty's to enter?

A few days ago I went ahead and...
  • Made seven 1/2 pints filled a little over halfway each. I don't plan on innoculating them all at once. I'm shooting to stagger that process over several weeks and have some on hand if I run into contams. 
  • They're regular mouth with the gray leakproof Ball lids
  • I PCd with the lids loose and covered with foil
  • Let the PC cool all the way with the weight still on figuring that would maybe decrease the amount of air coming in as it cooled, yet be enough to even the pressure without it being a big swoosh when opening
  • Took them out and tightened the lids

... with the intention of cracking the lids to inoculate with your Poke method, once colonized and if it looks clean pour on a few grain jars. If they take and look clean, use those as masters for G2G. Once I get over the jitters of doing something new I'll move to innoculating all grain jars with LC... ??? Anyway, that's what I've come up with so far... having a hard time wrapping my head around getting and maintaining a momentum...

Another question... I can't keep taking a clone, putting it to agar, Poking it, pour on grain, harvest, repeat indefinitely can I? Doesn't that age the line (sorry if I'm not using the right shroom terminology... new to this)? I mean don't I have to gather spores and start fresh at some point in order to restart the aging/senescence process?


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Re: The Biopsy Method: A different way of doing LC. [Re: Threads2theSoul]
    #27319188 - 05/22/21 09:53 PM (4 months, 27 days ago)

Quote:

'contaminated air cannot travel through obscure pathways in all circumstances




I don't know if they were obscured or illuminated, but I've had jars contaminate that were never opened using only unmodified threaded plastic lids. And the circumstances was a place with mold, lots of mold.

Also not sure if Pasteur was referring to threaded plastic lids when he was talking about those obscure pathways. Not all quotes and stuff you find on the internet applies to cult.

If you took jars out while hot and didn't screw down the unmodified lids completely, the jars could perfectly suck contams in that could survive when the jars start to cool down. But if you closed them immediately they can create a vacuum due to the hot contents inside, and they could suck air in next time you open them.

I'm an individual with my own set of experiences and preferences, by no means what I'm saying is set in stone. So you can take or leave what I say and just try it yourself, maybe you like it and it doesn't give you problems.
There are things that may work under certain circumstances and you may never encounter an issue with them even if they're not 100% reliable under different circumstances.

For me, properly filtering LC lids is reliable and really works in all circumstances. I don't have to worry about cracking lids before the cycle, tightening them after, vacuums or jars sucking in air. I want none of that.
Do whatever feels right to you, experiment and judge for yourself.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
LAGM '21

PHENO HUNTERS


Edited by Josex (05/22/21 10:57 PM)


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