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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC [Re: A.k.a]
    #27134096 - 01/07/21 09:11 PM (3 years, 2 months ago)

Yup, it all goes fast once you start seeing the first little puffs. 2 or 3 swirls a day to keep them going.

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OfflineJarhead3521S
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Re: The Biopsy Method: A different way of doing LC [Re: Josex]
    #27134108 - 01/07/21 09:14 PM (3 years, 2 months ago)

Gotcha!  Yah I’ve been swinging into my shop when I get home to check plates and give em a lil swirly swirl......I’m diggin it though.  Just put one of the huatla plates and the PE plate that I poked to grains while another 10 jars are in the PC so I can put the other huatla and Malabar plate to those jars.


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OfflineTankie_J
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Re: Josex' Poke: A different way of doing LC. [Re: Josex]
    #27138736 - 01/09/21 11:02 PM (3 years, 2 months ago)

:cheers:

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OfflineJarhead3521S
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Re: Josex' Poke: A different way of doing LC. [Re: Tankie_J]
    #27139397 - 01/10/21 10:03 AM (3 years, 2 months ago)

My PE is thriving :rockon:


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OfflineJarhead3521S
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Re: Josex' Poke: A different way of doing LC. [Re: Jarhead3521]
    #27150037 - 01/15/21 04:23 PM (3 years, 2 months ago)



I hate asking these kinds of questions, but it’s my first go at LC so hell with it.  At what point is this “done and ready”


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Offline9chis
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Re: The Biopsy Method: A different way of doing LC. [Re: Jarhead3521]
    #27150069 - 01/15/21 04:38 PM (3 years, 2 months ago)

: bow2:

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InvisibleAyePlus
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Re: Josex' Poke: A different way of doing LC. [Re: Jarhead3521]
    #27150105 - 01/15/21 04:49 PM (3 years, 2 months ago)

Quote:

Jarhead3521 said:


I hate asking these kinds of questions, but it’s my first go at LC so hell with it.  At what point is this “done and ready”




That one is ready:grampofapproval:

Send it


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OfflineJarhead3521S
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Re: Josex' Poke: A different way of doing LC. [Re: AyePlus]
    #27150122 - 01/15/21 04:57 PM (3 years, 2 months ago)

Word! Looks like I’m cooking grains in the morning instead of hunting!


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OnlineA.k.aM
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Re: Josex' Poke: A different way of doing LC. [Re: Jarhead3521]
    #27150198 - 01/15/21 05:35 PM (3 years, 2 months ago)

Yep swirl that thing around and it’ll be loaded with myc bits.


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OfflineEdmunter
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Re: Josex' Poke: A different way of doing LC. [Re: A.k.a]
    #27155323 - 01/18/21 12:18 PM (3 years, 2 months ago)

Holy shit this TEK makes so much sense......  On it......will report back later.....  :kthxbye:

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Offlinegt40
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Re: The Biopsy Method: A different way of doing LC. [Re: Edmunter]
    #27155550 - 01/18/21 02:00 PM (3 years, 2 months ago)

Thanks Josex, it's truly LC gamechanger. What do you guys do with really dense myc grow inside LC? Sometimes it's superhard to break it. I use the biggest needle, may pieces of glass inside can help?

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OfflineEdmunter
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Re: The Biopsy Method: A different way of doing LC. [Re: gt40]
    #27155557 - 01/18/21 02:03 PM (3 years, 2 months ago)

Quote:

gt40 said:
Thanks Josex, it's truly LC gamechanger. What do you guys do with really dense myc grow inside LC? Sometimes it's superhard to break it. I use the biggest needle, may pieces of glass inside can help?




Some people use marbles

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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: Edmunter]
    #27155588 - 01/18/21 02:18 PM (3 years, 2 months ago)

:whathesaid:

I also used to use blender balls like in the pics:


Here you can get some for cheap. Just swirling gently gets the myc nice and shreded with these balls.

Even without anything to help you break the myc, swirling several times a day helps a lot to prevent big clumps from forming. If you're pouring your LC's, clumping is not so much of an issue, though.

You could also buy a cheap magnetic stirrer and some stir bars if you're so inclined.

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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27169431 - 01/25/21 04:21 PM (3 years, 2 months ago)

I'm looking to do LC so I can inoculate Mudafuka bottles via pouring. I'm trying to avoid modded lids and aspirating LC and just rely on media bottles and pouring to inoculate the Mudafuka substrate bottles.

Is it a crazy idea to test the LC made via poke biopsy by pouring to a single small grain jar prior to inoculating my bottles? Does the risk of contaminating the LC by opening, pouring and closing again to perform the test outweigh the benefits?

Or would I better be off testing the original source culture by doing the poke and squirt to a clean plate prior to making my broth as suggested in this tek?

Or not testing at all, and taking the risk assuming the the LC looks clear and clean and pouring to my bottles anyway?

Edited by bongoman (01/25/21 04:27 PM)

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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: bongoman]
    #27170250 - 01/26/21 04:37 AM (3 years, 2 months ago)

I'm truly bad at making contaminated LC, barely know what it is, so I'm spoiled and never feel I have to test it. Bullshit aside, the procedure for the poke is very foolproof and safe so you can rely on your good judgement if you're sure you're working with a clean culture. Test it if that's what you like to do and you aren't sure or don't feel confident yet.

Quote:

Or would I better be off testing the original source culture by doing the poke and squirt to a clean plate prior to making my broth as suggested in this tek?





I wrote that years ago but today I'd rather tell you to make an LC and test it of you're unsure. If you don't have a modified lid for aspirating, the best way to test an LC is reaching with a flamed inoc loop and putting it to agar.

Also, bare in mind opening the LC jar is a risk in itself. You could introduce contams in what could have been a clean LC before.

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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27172076 - 01/27/21 12:42 AM (3 years, 1 month ago)

Quote:

Josex said:
I'm truly bad at making contaminated LC, barely know what it is, so I'm spoiled and never feel I have to test it. Bullshit aside, the procedure for the poke is very foolproof and safe so you can rely on your good judgement if you're sure you're working with a clean culture. Test it if that's what you like to do and you aren't sure or don't feel confident yet.

Quote:

Or would I better be off testing the original source culture by doing the poke and squirt to a clean plate prior to making my broth as suggested in this tek?





I wrote that years ago but today I'd rather tell you to make an LC and test it of you're unsure. If you don't have a modified lid for aspirating, the best way to test an LC is reaching with a flamed inoc loop and putting it to agar.

Also, bare in mind opening the LC jar is a risk in itself. You could introduce contams in what could have been a clean LC before.





Yo Josex!!!

So a couple of questions.

Do you ever miss grabbing a sample during the stab?  I think I did

How do you test, I mean details if possible

I loaded my syringes in front of the Flow hood but without a filter it draws in air from outside.  Has this ever caused a problem?

Do you create a clean jar and then G2G?

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OnlineA.k.aM
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Re: The Biopsy Method: A different way of doing LC. [Re: Edmunter]
    #27172114 - 01/27/21 01:47 AM (3 years, 1 month ago)

My first shot at this I made three and only one got myc in it so it’s definitely possible.


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InvisibleJosex
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Re: The Biopsy Method: A different way of doing LC. [Re: A.k.a]
    #27172148 - 01/27/21 02:58 AM (3 years, 1 month ago)

Yea as aka said you can miss, it happens.

To avoid that I make sure I poke established myc on the plate so I avoid myc on the edges or spots that look more wispy. Also, poking the culture more than once before squirting the tissue def helps, there's no issues with that if your plate is clean. How long has been anyway? Sometimes the sample takes time if it's particularly small (can take up to six days to recover in rare cases).

I rarely ever test Ed. Currently I aspirate into syringes and the few times I did test I just put a drop to agar. When I used to pour (from a plain quart jar without modifications for aspirating) and wanted to test I got a red hot long loop in the jar and streaked it on agar.


Quote:

I loaded my syringes in front of the Flow hood but without a filter it draws in air from outside.  Has this ever caused a problem?




All my LC lids are properly filtered, not a good idea getting air in from the outside, but likely you can get away with it in the flow.

Quote:


Do you create a clean jar and then G2G?




I find g2g redundant in my situation when I can easily make way more LC that I could possibly use. Also, making axenic LC is easy, axenic spawn not so much...

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Re: The Biopsy Method: A different way of doing LC. [Re: Josex]
    #27172152 - 01/27/21 03:02 AM (3 years, 1 month ago)

I finally read this all the way through and Boy oh Boy is it entertaining. Great write up Josex!

  I am going to order some SFD's and give this ago. None of my lids are currently compatible with this method.

Question, can you poke to plate? Why wouldn't this work for agar transfers?

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Re: The Biopsy Method: A different way of doing LC. [Re: Sockadin]
    #27172160 - 01/27/21 03:14 AM (3 years, 1 month ago)

It does. He’s got another poke method for cleaning bacterial plates.


Poking thicker myc multiple times and/or twisting the needle while it’s in the agar has definitely increased my success rate. There’s a little bit of a learning curve but it’s easy.


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Edited by A.k.a (01/27/21 03:16 AM)

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