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OfflineSold Out Online
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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27068604 - 12/02/20 06:37 PM (10 months, 7 days ago)

I split 500ml across two jars, thinking I didn’t want to have potentially contaminated leftovers.

But... then this morning I poke inoculated two quart jars with 500 ml each using 1g of LME each... so now I’ll just have to deal with it. Going to need a taller SAB.  :freewilly:  :freewilly:  :freewilly:


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Re: The Biopsy Method: A different way of doing LC. [Re: Sold Out Online]
    #27069354 - 12/03/20 08:18 AM (10 months, 6 days ago)

500ml per jar takes way too long, that ain't wise anymore bro.
If you split it between 2 jars like you did first, you'll have the same amount of LC in less than half the time.

You inoculated those 500ml LC just recently so I'd personally throw them down the toilet right now unless you're willing to wait roughly one month for them to be done. I'd make new ones with no more than 250ml and if you want them ready even faster 200ml is where it's at.


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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]
    #27069372 - 12/03/20 08:47 AM (10 months, 6 days ago)

I’ve got no shortage of jars and am in no rush so I’ll just leave them doing their thing and make up some more. In the meantime they’ll be educational to watch.

Royally fucked my first grow so I’m just trying to nail down a bunch of clean cultures so I can rapid-fire try to get something to fruit without having to go back to the online spore dealers.


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Re: The Biopsy Method: A different way of doing LC. [Re: Sold Out Online]
    #27077483 - 12/07/20 07:53 PM (10 months, 2 days ago)

I had almost given up. My two 500 ml jars of water-clear LME broth had no visible growth. I had no faith my pokes had picked anything up. After inoculating 4 quarts with this nasty stuff:



I had just a few drops left and I thought I might as well use it on my pristine broth. Then I glanced up at my shelf and saw this little guy floating around...



The other jar has one that looks the same. The poke works, just gotta have faith.

Before pouring my dirt nasty 3% LME to grains I dribbled a bit into these four rye grain broths I made, 1/2 diluted with water.



I’m glad I found this tek!


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Re: The Biopsy Method: A different way of doing LC. [Re: Sold Out Online]
    #27078779 - 12/08/20 03:17 PM (10 months, 1 day ago)

Jalisco at the start of day 13. Took a minute to get going but it’s really blown up the last couple days.



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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27079043 - 12/08/20 06:40 PM (10 months, 1 day ago)

Quote:

Josex said:
500ml per jar takes way too long, that ain't wise anymore bro.
If you split it between 2 jars like you did first, you'll have the same amount of LC in less than half the time.

You inoculated those 500ml LC just recently so I'd personally throw them down the toilet right now unless you're willing to wait roughly one month for them to be done. I'd make new ones with no more than 250ml and if you want them ready even faster 200ml is where it's at.




Your so right on smaller amounts.

Blue Helix Lc recipe:
1000 ml. RO water.
20 grams LME. (I used Amber Malt Extract)
20 grams Dextrose (corn sugar)

https://www.shroomery.org/forums/showflat.php/Number/27010409#27010409

140 ml. broth per jar.
7 salsa jars.



7 days growth. I used 5ml. Lc. to expand each of my 7 jars...



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Re: The Biopsy Method: A different way of doing LC. [Re: micelio]
    #27079053 - 12/08/20 06:50 PM (10 months, 1 day ago)

7 days, now we're talking, lc to lc is fast. That's a lot of nutes tho. I could show you some really thick stirred and non-stirred LC'S at 0.2% strength.
Incidentally, I've been postponing it but I promise any day soon I'll do a thread on how I do LC's to be ready in 6-7 days from biopsy, it is certainly possible.


--------------------
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]
    #27079106 - 12/08/20 07:22 PM (10 months, 1 day ago)

Quote:

Josex said:
7 days, now we're talking, lc to lc is fast. That's a lot of nutes tho. I could show you some really thick stirred and non-stirred LC'S at 0.2% strength.
Incidentally, I've been postponing it but I promise any day soon I'll do a thread on how I do LC's to be ready in 6-7 days from biopsy, it is certainly possible.




I know It's allot of nutes..
I don't know what got into me..
It  was one of those moments that I said to myself..,  Okay I'll bite..

I have never had that fast of growth.  Thank you Blue Helix..
It's fun to experiment and get away from what's suppose to be the normal...

Josex.., I'm looking forward to your thread on 6-7 day from a biopsy..


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Re: The Biopsy Method: A different way of doing LC. [Re: micelio]
    #27082838 - 12/10/20 08:48 PM (9 months, 30 days ago)

First time trying LC and have no idea what I'm looking at here but it doesn't look good:





2/3 jars showing blobby growth like this about a week after poking a TOC culture. One jar showing no growth at all.

Gonna give it a stir cause I haven't touched it since I inoculated but do I even have cube myc here? Don't think I've really seen anything like this in the thread so far.


Thanks for the tek Josex!


Edit:

Just stirred these guys by hand. Looks a bit more normal now.


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:chems:


Edited by Lenz (12/10/20 09:02 PM)


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Re: The Biopsy Method: A different way of doing LC. [Re: Lenz]
    #27083254 - 12/11/20 01:52 AM (9 months, 29 days ago)

Those first pics with the bubble thing are weird man never seen that.


Unfortunately those look like bacteria colonies.


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Re: The Biopsy Method: A different way of doing LC. [Re: A.k.a]
    #27083405 - 12/11/20 05:34 AM (9 months, 29 days ago)

Quote:

Lens said:


Gonna give it a stir cause I haven't touched it since I inoculated but do I even have cube myc here? Don't think I've really seen anything like this in the thread so far.




Those blobs is what happens when you don't swirl the LC at all, you should have started swirling those days ago to break it up and speed things up.
It's perfectly normal for that to happen tho and they're looking good, no need to worry.

Do a search on this thread using the key word 'jellyfish' if you want to see instances of this. Your jellyfishes just happen to be the phatest in this thread, a pity you don't get to win a prize 🏆.


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LC's are easy
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Clean yo shit
LAGM '21

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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27083574 - 12/11/20 10:06 AM (9 months, 29 days ago)


Hey everyone im diving into LC with this tek for the first time and did not yet feel like being nasty as fuck :hehehe: so I made 3 LME broths and I'm guessing I did it wrong. I don't think I cooked it enough pre PC or something because there's just LME floating around in there that settles at the bottom if not stirred as you can see in the pic.

I used the 1g to 500ml recipe for 1 of them and that one isn't so cloudy but still just has some LME floating in it but the two I packed on more nutes have lots of LME just a'floatin. This is normal?

I pressure cooked 15psi for 30 minutes so I'm probably still going to use these. I don't see why extra LME floating would hurt just less clarity and extra nasty as fuck :rockon:. I guess I was expecting just a clear liquid indistinguishable from water but the LME doesn't seem to have cooked in. :shrug:

Waiting for a response then I'm gonna get these things poked and pregnant thanks for the tek. I hope this goes well :super:

Edit: Reading through some pages here I'm probably going to just re-do it I did mess it up somehow. I want the crystal clear water like broth my first time there's a bit too much LME in mine.
Quick poke question. Can I use one needle to poke and noc the 3 jars flame sterlizing in between? Or should I use three separate syringes? That seems like it'd be a waste of syringes one per session but I might be misunderstanding
And also does anyone use this poke to do agar to agar transfers? I feel like that'd be useful but I also see the benefit of BRF, just need to get into it.


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


:sporedrop:Pho's Mushi Aventures:sporedrop: : First Agar Journey:bow2:  Pho Conquors Agar

:buddha: LAGM2021 :buddha2:


Edited by Ababyphoenix (12/11/20 10:23 AM)


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Re: The Biopsy Method: A different way of doing LC. [Re: Ababyphoenix]
    #27083613 - 12/11/20 10:44 AM (9 months, 29 days ago)

as long as you flame sterilize though one syringe is all you need

In regards to A2A transfers it depends on what you are looking for. You will get clean mycelium, thats for sure. But if you are looking to isolate individual sectors or clean a moldy plate its a nono. Remember that with A2A transfers you put the piece in the center and let it grow out, it doesnt work the same way when you poke and squirt


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Re: The Biopsy Method: A different way of doing LC. [Re: Ababyphoenix]
    #27083690 - 12/11/20 11:44 AM (9 months, 29 days ago)

If you have doubts about your syringe..

Boil some water in a small pot and cycle the boiled water in and out of your syringe for a few minutes...

Don't forget to flame the needle..

This is excellent info. on cloudy Lc. It's a must read...

https://www.shroomery.org/forums/showflat.php/Number/26977498/fpart/1/vc/1


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Re: The Biopsy Method: A different way of doing LC. [Re: Sankhara]
    #27083719 - 12/11/20 11:59 AM (9 months, 29 days ago)

Quote:

Sankhara said:

In regards to A2A transfers it depends on what you are looking for. You will get clean mycelium, thats for sure. But if you are looking to isolate individual sectors or clean a moldy plate its a nono. Remember that with A2A transfers you put the piece in the center and let it grow out, it doesnt work the same way when you poke and squirt




Well yes of course it doesn't what I've been doing is poke and inject more of the center of the agar dish. shoot that sum 'bish up if you get. It worked alright ish the wider gauge needle just too much water. Trying again with the 21G and I don't see how it wouldn't work. :shrug:
Even with the poke and squirt trials I did on agar i still got growth on the center just don't toss ur plate around like its your bitch before it grows and grabs the agar. Condensation sliding around on the surface is common for petris anyway as long as u have good sterile technique it doesn't hurt a thing.

Edit (some proof):
These were with the poke and squirt onto the surface. Still waiting on 21g Transfers with the poke n inject to show growth, but its been one day so :cheers:
It actually looks like it started a few colonies as it spread so I'm not complaining. Especially since I don't see any satellites from using a needle over top the dish


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


:sporedrop:Pho's Mushi Aventures:sporedrop: : First Agar Journey:bow2:  Pho Conquors Agar

:buddha: LAGM2021 :buddha2:


Edited by Ababyphoenix (12/11/20 12:08 PM)


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Re: The Biopsy Method: A different way of doing LC. [Re: Ababyphoenix]
    #27083751 - 12/11/20 12:14 PM (9 months, 29 days ago)

Nice! Thats cool. when doing transfers like that, do you squirt or just put a tiny drop?

I get a feeling that a drop or no drop at all (striking plate with needle) should work just fine, never done it though.


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Re: The Biopsy Method: A different way of doing LC. [Re: Sankhara]
    #27083875 - 12/11/20 01:40 PM (9 months, 29 days ago)

I did it both ways. I squirted as little as I could with a wide gauge needle (so a decent amount of water) onto the surface and that's whats pictured there. Waiting on the inject and squirt now. Should come out more like a normal transfer me thinks


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Re: The Biopsy Method: A different way of doing LC. [Re: Ababyphoenix]
    #27105442 - 12/24/20 10:46 AM (9 months, 16 days ago)

Lil late to the party but thanks for the tek!
I made 2 LC's yesterday with this method and will keep you guys updated.
The agar dish I poked seem to be recovering nicely already with the myc starting to grow over the soot from the needle.
The anxiousness this tek describes like 'no way in hell will this work' immediately kicked in when working with such small tissue. Can't wait to be pleasantly surprised. At the same time I have very little experience with agar so if this turns out to be a dud it's most likely on me knocking my SAB around a bit because it's smaller than I want it to be lol.


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Re: The Biopsy Method: A different way of doing LC. [Re: CorteX_]
    #27105589 - 12/24/20 12:44 PM (9 months, 16 days ago)

Have faith, child.

:pope:


--------------------
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]
    #27106993 - 12/25/20 09:26 AM (9 months, 15 days ago)

Using the poke I’ve successfully colonized these two jars of 0.2% LME, 500ml each:



And when I was pouring my last LC (also poke tek) to grain I LC2LC’d four rye broth jars...



One looks fucked. Flattens out like that even after vigorous shakes.

Now I need to figure out how the hell I’m ever going to get through 1600ml of GT LC.


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