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InvisibleDrboomer
The lord magnificent
I'm a teapot

Registered: 09/22/19
Posts: 566
Re: The Biopsy Method: A different way of doing LC. [Re: gone-pear-shaped]
    #27347533 - 06/14/21 06:42 AM (3 months, 2 days ago)



Just finished a dozen of these. They are pint jars joined to quarter pint jars. I epoxied the rings togather then put two layers of epoxy around the outside. It's pretty sturdy I tried beating one apart with a hammer and chisel and destroyed the ring before I got them apart.


I cut the center out of one jars lid so both sides had gaskets. Drilled two 1/4 holes in the pint jars lid. 1 hole gets polyfil the other 1/4 28tpi polypropylene female luer lock bulkhead with a 1/8 hose Barb on top of a 1/4x 3/4 flat washer all rtvd into place and held on with a 1/4x28tpi stainless nut. After this picture I drilled a 1/4 hole in the top of the small jar then epoxied down a large piece of an sfd, waited for it to dry then expoxied around it's perimeter it's stuck on pretty well.

The bulkhead and tubing came from Amazon, everything else from my local big box hardware store.

I'm hoping to make 200 to 225 ml of lc at a time. 9 of these fit into my 21qt old presto at a time and a small stainless steel box fits on top with foil wrapped syringes. I'm going to make a half gallon one to pour lc from ovoids all over wood chips outside by my favorite hiking path. I know it will take forever to colonize that volume but it is just a fun project.


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OfflineThreads2theSoul
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Re: The Biopsy Method: A different way of doing LC. *DELETED* [Re: Drboomer]
    #27349967 - 06/15/21 11:36 PM (3 months, 11 hours ago)

Post deleted by Threads2theSoul

Reason for deletion: found the answer ... "Always use freshly sterilized syringes. Don't cut corners here and never store syringes for later use."


--------------------
Granny's Awesome LAGM21


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InvisibleVikingWizard
Mycologist


Registered: 06/16/21
Posts: 23
Re: The Biopsy Method: A different way of doing LC. [Re: Threads2theSoul]
    #27350052 - 06/16/21 01:53 AM (3 months, 9 hours ago)

Great informative post thanks for sharing.


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Invisiblepablokabute
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Re: The Biopsy Method: A different way of doing LC. [Re: VikingWizard]
    #27354730 - 06/19/21 05:08 AM (2 months, 28 days ago)

Is it really close to impossible to successfully do LC? Can the smallest tissue outrun bacterial contamination in plates? Thanks.


--------------------

Fermented Mushrooms!!
--- https://www.shroomery.org/forums/showflat.php/Number/23378638/fpart/1/vc/1

'The second seal: “All CONTAMINATED things and events are unsatisfactory.”'



"I envy you. You North Americans are very lucky. You are fighting the most important fight of all - you live in THE HEART OF THE BEAST."

--Anonymous Guerilla, or is he..


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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: pablokabute]
    #27354864 - 06/19/21 08:57 AM (2 months, 28 days ago)

You don't want to outrun contamination on agar and the term "outrun" is inconceivable for LC, as any CFU's present quickly and exponentially reproduce and multiply in the liquid, rendering it useless.

You want to aim for axenic cultures on agar and more so in LC. It's not only possible but fairly easy to the point where I can count with one hand the times I've failed with LC, and I've done a lot. Hope you can fix your issues soon man, you can shoot any questions you have.


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

PHENO HUNTERS


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Invisiblepablokabute
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27354872 - 06/19/21 09:14 AM (2 months, 28 days ago)

Quote:

Josex said:
You don't want to outrun contamination on agar and the term "outrun" is inconceivable for LC, as any CFU's present quickly and exponentially reproduce and multiply in the liquid, rendering it useless.

You want to aim for axenic cultures on agar and more so in LC. It's not only possible but fairly easy to the point where I can count with one hand the times I've failed with LC, and I've done a lot. Hope you can fix your issues soon man, you can shoot any questions you have.





Thanks for the reply.

There ya go. I learned a new term. Axenic. I was wondering of its possible to clean bacterial plates without resorting to antibiotic media?

I will try again in a few days or weeks and post back. Thanks for the neat writeup, my man. Ive attempted LC a couple of times actually. It almost appears impossible to accomplish with just a SAB but could very well be my technique too, I havent saved for a hood yet. 

Will try this with oysters, too. Heheh


--------------------

Fermented Mushrooms!!
--- https://www.shroomery.org/forums/showflat.php/Number/23378638/fpart/1/vc/1

'The second seal: “All CONTAMINATED things and events are unsatisfactory.”'



"I envy you. You North Americans are very lucky. You are fighting the most important fight of all - you live in THE HEART OF THE BEAST."

--Anonymous Guerilla, or is he..


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OfflineA.k.a
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Re: The Biopsy Method: A different way of doing LC. [Re: pablokabute]
    #27354878 - 06/19/21 09:17 AM (2 months, 28 days ago)

Lc is pretty much the same difficulty level as doing clean plate transfers. If you can consistently make clean plate transfers you should be fine. The only difference is that a slightly dirty culture might not be detectable on a plate but will absolutely explode in an lc.


Then using the poke method makes things even easier.


--------------------
LAGM2020


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Invisiblepablokabute
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Re: The Biopsy Method: A different way of doing LC. [Re: A.k.a]
    #27354883 - 06/19/21 09:21 AM (2 months, 28 days ago)

Quote:

A.k.a said:
Lc is pretty much the same difficulty level as doing clean plate transfers. If you can consistently make clean plate transfers you should be fine. The only difference is that a slightly dirty culture might not be detectable on a plate but will absolutely explode in an lc.


Then using the poke method makes things even easier.





Yea man. It appears the lesser amount of transfer material = slightly better chance of clean culture. Imma grab me a bunch of 21Gs when at the drugstore.


--------------------

Fermented Mushrooms!!
--- https://www.shroomery.org/forums/showflat.php/Number/23378638/fpart/1/vc/1

'The second seal: “All CONTAMINATED things and events are unsatisfactory.”'



"I envy you. You North Americans are very lucky. You are fighting the most important fight of all - you live in THE HEART OF THE BEAST."

--Anonymous Guerilla, or is he..


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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: pablokabute]
    #27355064 - 06/19/21 12:44 PM (2 months, 27 days ago)

I personally dislike using the poke to do agar transfers, a scalpel is the correct tool for the job.
You can perfectly work with a agar in a SAB, using a scalpel for transfers with a really high success rate. Same with LC, you can do it a SAB and I came up with this method at the time because it makes it all more foolproof if you're working in a SAB

If you think you have a dirty culture you can try my puck method, I've cleaned stuff that way many times. And if you feel more comfortable working with the poke than a scalpel, then pucks are also good for that, as the substrate will absorb the liquid when you squirt.

I never had a FH myself, people that tell you that you can't have a high success rate without a hood are spreading misinformation. Hell I work open air with just a bunsen burner and I never see contams :shrug:


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

PHENO HUNTERS


Edited by Josex (06/19/21 01:20 PM)


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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27355072 - 06/19/21 01:00 PM (2 months, 27 days ago)

Have to add that I've yet to see someone who sucked at growing before owning a FH that didn't keep sucking after having it.


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

PHENO HUNTERS


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OfflineSpaceBabyS
alchemist
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Registered: 11/01/20
Posts: 951
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27360648 - 06/23/21 10:09 PM (2 months, 23 days ago)

06.22

:hatsoff:
Josex' Poke



Never did LC before, so the q is...is it ready?
Since it's a narrow mouth pint with about 150 ml in it, I can just split it among 10 jars?

agar wedge



--------------------
SpaceBaby
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OfflineBrownBear
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Re: Josex' Poke: A different way of doing LC. [Re: SpaceBaby]
    #27360676 - 06/23/21 10:26 PM (2 months, 23 days ago)

Those lc's look nice and ready. Especially that first one. Its nice and thick the way I like it.


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OfflineSpaceBabyS
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Re: Josex' Poke: A different way of doing LC. [Re: BrownBear]
    #27360907 - 06/24/21 12:30 AM (2 months, 23 days ago)

Quote:

Iambrownbear said:
Those lc's look nice and ready. Especially that first one. Its nice and thick the way I like it.




Thanks, BB. That is my very first LC. Josex' Poke worked sweetly. I'm going to get some oats going and pour ten jars. I had 4 1/2-pints Poked and this is the only on that not only looks good, it doesn't looked horrific. Pics of those to follow—I'm down for the night after three days of hella headaches. The media bottle [I have two other LCs going, the total is three] became a possibilty when I PCd 6 bottles of agar and then discovered I' d put everything except the agar in three. :rofl:

That said, I'd rather lose individual, or even all 10, jars of grain than indubitably contaminate the 1/2-pint mason jar. If I get anything I can do Pasty's Ziplock or just a shoebox. Calculated risk; I don't wanna start a new LC until I do a couple of transfers of my new Malabar clone.

Fuckin long-winded, all-about-me, thanks.


--------------------
SpaceBaby
SPACEBABY's LAZY FUCKER SHOEBOX LINING TEK

tla Reine de la Maison:

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InvisibleJosex
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Re: Josex' Poke: A different way of doing LC. [Re: SpaceBaby]
    #27360927 - 06/24/21 12:51 AM (2 months, 23 days ago)

Nice, Space. Keep that shit up, loving everything you do but the oats, as you know haha


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

PHENO HUNTERS


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OfflineSpaceBabyS
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27360942 - 06/24/21 01:06 AM (2 months, 23 days ago)

Quote:

Josex said:
Nice, Space. Keep that shit up, loving everything you do but the oats, as you know haha




Thanks Josex, you been swell to me and I'm :bow2:

Yeah. But I just got hella kitted-out in the hobby. Start-up costs are a bitch. :whiteflag: I still got oats; I'm using oats till at least I ain't got any. After that I might try wheat or millet. That said, aside from 2 nasty contam jars [so far:rolleyes:] this Huatla looked good. It's probably the oats is why they were a total cunt to break and shake, dunno.

Like I said, in the last couple days I pulled 2.5-pounds wet out of them. And there are more than 2-dozen shoeboxes around here in various stages of colonizing/fruiting.


--------------------
SpaceBaby
SPACEBABY's LAZY FUCKER SHOEBOX LINING TEK

tla Reine de la Maison:

porn:


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Invisiblepablokabute
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Re: Josex' Poke: A different way of doing LC. [Re: SpaceBaby]
    #27361033 - 06/24/21 03:57 AM (2 months, 23 days ago)

Quote:

SpaceBaby said:
Quote:

Josex said:
Nice, Space. Keep that shit up, loving everything you do but the oats, as you know haha




Thanks Josex, you been swell to me and I'm :bow2:

Yeah. But I just got hella kitted-out in the hobby. Start-up costs are a bitch. :whiteflag: I still got oats; I'm using oats till at least I ain't got any. After that I might try wheat or millet. That said, aside from 2 nasty contam jars [so far:rolleyes:] this Huatla looked good. It's probably the oats is why they were a total cunt to break and shake, dunno.

Like I said, in the last couple days I pulled 2.5-pounds wet out of them. And there are more than 2-dozen shoeboxes around here in various stages of colonizing/fruiting.





What size shoebox and what tek are you following? :smile:


--------------------

Fermented Mushrooms!!
--- https://www.shroomery.org/forums/showflat.php/Number/23378638/fpart/1/vc/1

'The second seal: “All CONTAMINATED things and events are unsatisfactory.”'



"I envy you. You North Americans are very lucky. You are fighting the most important fight of all - you live in THE HEART OF THE BEAST."

--Anonymous Guerilla, or is he..


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Offlinerockyfungus
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Re: Josex' Poke: A different way of doing LC. [Re: pablokabute]
    #27361106 - 06/24/21 07:32 AM (2 months, 23 days ago)

Quote:

pablokabute said:


There ya go. I learned a new term. Axenic. I was wondering of its possible to clean bacterial plates without resorting to antibiotic media?





If you want to avoid antibiotics. Use the poke to isolate clean material, or growing some myc out on brf or grain and transferring that grain to new agar can work. Just takes a bit of patience.

Others use "agar sandwiches" or a warmer pour of the agar.

Sorry struggling to find links and I suck with words.


--------------------
Team frugal


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Offlinegone-pear-shaped
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Re: Josex' Poke: A different way of doing LC. [Re: rockyfungus]
    #27361108 - 06/24/21 07:37 AM (2 months, 23 days ago)

Quote:

rockyfungus said:Others use "agar sandwiches" or a warmer pour of the agar.
Sorry struggling to find links and I suck with words.



The latter is often called a "hot pour".


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OfflineSpaceBabyS
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Re: Josex' Poke: A different way of doing LC. [Re: pablokabute]
    #27361123 - 06/24/21 08:01 AM (2 months, 23 days ago)

Quote:

pablokabute said:
Quote:

SpaceBaby said:
Quote:

Josex said:
Nice, Space. Keep that shit up, loving everything you do but the oats, as you know haha




Thanks Josex, you been swell to me and I'm :bow2:

Yeah. But I just got hella kitted-out [NOT a kit, haha] in the hobby. Start-up costs are a bitch. :whiteflag: I still got oats; I'm using oats till at least I ain't got any. After that I might try wheat or millet. That said, aside from 2 nasty contam jars [so far:rolleyes:] this Huatla looked good. It's probably the oats is why they were a total cunt to break and shake, dunno.

Like I said, in the last couple days I pulled 2.5-pounds wet out of them. And there are more than 2-dozen shoeboxes around here in various stages of colonizing/fruiting.





What size shoebox and what tek are you following? :smile:






7-qt
Started out Bod's EAF Teks

Oats
Straight coir Bucket TEK
1 myco-qt spawn to ~2-quarts sub
Adjust ecosystem by examination
Harvest as close to veil-tear as possible

I have been cloning the 'best' fruits in terms of size/speed of growth so that I can get a sense of the strength. I will be learning Masters, Slaves, amd Slants soon. Will keep the BOTB.
Taking spore prints of some.

Also running one fahtser Ziplock Tek, it's colonizing far slower than faht says, but it's colonizing. am ever more proficient with agar, and transfers, and accept that failures are merely education.

I have two boxes that are waiting for me to pull fruits rn. Am twisting until canopies make 'scything the surface' possible. The fruits are popping up and tearing continuously at this point.

Spawning another myco-quart later today after 'work.'

Tried 3 g dry of my Malabar Sunday and loved it. Nice low Level Three, although that is only a generally information statement.

Enough data?

Since it's "JoPo" here's an appropriate "pic tax."



All the same B+ clone I named 'Superbad' because the baby was harvested out of a pf tek cake 9-days from introduction to the SGFC and before there was another pin on any of the three varietal cakes. Although, generally, RN all my Malabar is the quickest producer.

IFLM


--------------------
SpaceBaby
SPACEBABY's LAZY FUCKER SHOEBOX LINING TEK

tla Reine de la Maison:

porn:


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Invisiblenix21
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Re: Josex' Poke: A different way of doing LC. [Re: SpaceBaby]
    #27375943 - 07/06/21 08:54 AM (2 months, 11 days ago)

If i wanna do more then one LC with same needle and syringe should i sanitize syringe with alcohol between every needle flaming and poking?


--------------------


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