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InvisibleMr. Alien
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Registered: 01/14/14
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Re: The Biopsy Method: A different way of doing LC. [Re: Madcapper144]
    #26396493 - 12/22/19 10:17 PM (4 years, 3 months ago)

With fire? Is all metal. And what you mean after? you meant before i hope.

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Offlinevikingsc
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Re: The Biopsy Method: A different way of doing LC. [Re: Mr. Alien]
    #26397597 - 12/23/19 02:14 PM (4 years, 3 months ago)

Quote:

Mr. Alien said:
Would this work? IS exactly the same as a syringe but without water.

https://www.wpiinc.com/var-504639-reusable-rapid-punch-biopsy-kit

Here's the manual so you see what i mean.





Some people use a leather punch like these
https://www.amazon.ca/Stainless-Hollow-Leather-Cutter-0-5mm-5mm/dp/B01NBM6EKE/ref=sr_1_7?keywords=leather+punch+set&qid=1577135596&sr=8-7

I'm going to get a set to try it out

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InvisibleMr. Alien
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Posts: 6,290
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Re: The Biopsy Method: A different way of doing LC. [Re: vikingsc]
    #26397615 - 12/23/19 02:21 PM (4 years, 3 months ago)

Quote:

vikingsc said:
Quote:

Mr. Alien said:
Would this work? IS exactly the same as a syringe but without water.

https://www.wpiinc.com/var-504639-reusable-rapid-punch-biopsy-kit

Here's the manual so you see what i mean.





Some people use a leather punch like these
https://www.amazon.ca/Stainless-Hollow-Leather-Cutter-0-5mm-5mm/dp/B01NBM6EKE/ref=sr_1_7?keywords=leather+punch+set&qid=1577135596&sr=8-7

I'm going to get a set to try it out




But then how you expel/eject it with the leather punch?

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Offlinevikingsc
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Re: The Biopsy Method: A different way of doing LC. [Re: Mr. Alien]
    #26398467 - 12/24/19 12:49 AM (4 years, 3 months ago)

https://m.facebook.com/groups/967598903348873?view=permalink&id=2395817607193655

I assume you likely need to be part of the shroomery Facebook page to be able to view the link. That video she's using one of the larger size ones for transfers. But you can use the smaller ones obviously if you wanted to.

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InvisibleMr. Alien
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Registered: 01/14/14
Posts: 6,290
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Re: The Biopsy Method: A different way of doing LC. [Re: vikingsc]
    #26398930 - 12/24/19 09:55 AM (4 years, 3 months ago)

Quote:

vikingsc said:
https://m.facebook.com/groups/967598903348873?view=permalink&id=2395817607193655

I assume you likely need to be part of the shroomery Facebook page to be able to view the link. That video she's using one of the larger size ones for transfers. But you can use the smaller ones obviously if you wanted to.




Can't see it, i guess i need first to join, i just sent the group request.

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OfflineMadcapper144
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Re: The Biopsy Method: A different way of doing LC. [Re: Mr. Alien]
    #26399402 - 12/24/19 04:30 PM (4 years, 3 months ago)

I gotcha didn't realize you can just torch it cool!!


--------------------
Buy the ticket, take the ride!! :fearandloathing:

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OfflinePrietenul
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Registered: 11/22/19
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Re: The Biopsy Method: A different way of doing LC. [Re: Madcapper144]
    #26443826 - 01/20/20 08:20 PM (4 years, 2 months ago)

So I've read through the whole 56 pages and I still did not see any consensus about the least risky way for aspiring the LC into syringes. (I don't want to drill glass and buy turkey tubes and stuff, SHIPS are said to not be sterile and you would still have to tilt the jar,..). Pouring bags in my SAB is not an option.

As I want to grow Pans after Asuras TEK (120ml to inoc one sealed bag, and I have two bags) I need to fill four 60ml syringes from a quart jar that has ~250ml of colonized broth in it.

Last time I tried this in the SAB it was the biggest pain in the ass, since starting this hobby, and of course I got contams in the end. Unscrew the lid and then fight with the tilted jar and syringe to get 60ml of myc into four syringes is shitty as hell.

The method I'm planning to try now is the following (taken mostly from this thread):

1.)
Fill my 60ml syringes and as well as a pint jar with freshly mixed LC broth.
2.)
PC all of it (syringes will lose some of the broth that is in them)
3.)
Poke the clean agar culture with syringe (that holds sterile broth)
4.)
Pull needed additional amount of broth from the pint jar into each syringe. This pulls the culture into the syringe and fills it up back to 60ml. (should be easy and fast even with short needles)
5.)
Cap syringes, seal or put in ziplock-bags and let grow out inside them
6.)
Flame sterilize needle and inoculate the bags in SAB


I think this procedure has the least contam vectors because the broth in the syringe is sterile and I don't have to open and pull everything from colonized LC jars. (biggest contam vector, IMHO)

The only objection I've read so far was about the visibility of the LC. For me this is totally unimportant, because a) I wouldn't know for sure, if I had contams in the jar by looking at it anyway, and thus b) I'd have to rely on clean procedures no matter what. So if it's clean it's clean and if not, it's not, no matter if I can see it perfectly clear. If I weren't sure about cleanliness I'd have to test on agar anyway.

Also, let's not forget, that a syringe is not opaque or so. You can still view the stuff pretty well. So if contams are that bad to be visible, you can see it through the syringe, too.

So before I start, I would really appreciate the opinions of other more seasoned cultivators or even someone who tested something like this.

Possibly there is another compelling reason not to do this or maybe someone found an even better way by now.

Thanks in advance for any input.

Edit: Also, wouldn't a syringe colonize much faster due to the smaller amount of liquid to be colonized?

Edited by Prietenul (01/21/20 08:02 AM)

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InvisibleComebackKid
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Re: The Biopsy Method: A different way of doing LC. [Re: Prietenul]
    #26444335 - 01/21/20 06:16 AM (4 years, 2 months ago)

There are no nutrients in the syringe. Just water.

You poke the sterile water syringe into the agar and then shoot the piece of agar into the jar with nutrients.

The LC is left to colonize and then when it's ready, you open the jar lid and pour the LC into your grains or whatever


Syringe is only used to inoculate the LC jar


--------------------
:amanita2: Substrate surface conditions / Monotub prep and care :sporedrop:

Look around you... Everything you see exists inside the mind.
Consciousness, the awareness that is experiencing this mind,
is peering in from outside the universe.

Our individual experiences are all part of the universe's experience of itself

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OfflineThe Mycologist
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Re: The Biopsy Method: A different way of doing LC. [Re: ComebackKid]
    #26444364 - 01/21/20 06:51 AM (4 years, 2 months ago)

"Poke once, poke fast, poke smart..."


--------------------
"That you are here—that life exists, and identity;
That the powerful play goes on, and you will contribute a verse.”
― Walt Whitman, Leaves of Grass
:acidfire::tmckenna:

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OfflinePrietenul
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Registered: 11/22/19
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Re: The Biopsy Method: A different way of doing LC. [Re: ComebackKid]
    #26444376 - 01/21/20 07:08 AM (4 years, 2 months ago)

Quote:

ComebackKid said:
There are no nutrients in the syringe. Just water.




If I put nutes (LC broth) inside it, there are nutes in the syringe. :lol: You read my post, didn't you?

Quote:


You poke the sterile water syringe into the agar and then shoot the piece of agar into the jar with nutrients.

The LC is left to colonize and then when it's ready, you open the jar lid and pour the LC into your grains or whatever


Syringe is only used to inoculate the LC jar




I guess you must have just skimmed over my post. Not a problem. Would be nice if you read it again and give me your opinion.

Edit: Ah it's possible that I wrote too clumsy. I meant, that I put broth in the syringe and the pint jar, before PCing. My English can be improved...and I should go to sleep :lol:

Edited by Prietenul (01/21/20 07:12 AM)

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OfflinePrietenul
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Re: The Biopsy Method: A different way of doing LC. [Re: The Mycologist]
    #26445615 - 01/22/20 04:39 AM (4 years, 2 months ago)

Quote:

The Mycologist said:
"Poke once, poke fast, poke smart..."




What do you mean by that?

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InvisibleComebackKid
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Re: The Biopsy Method: A different way of doing LC. [Re: Prietenul]
    #26445684 - 01/22/20 06:18 AM (4 years, 2 months ago)

Poke once:
When you do the poke sometimes its tempting to poke again because it doesnt look or feel like you even picked up any myc... don't second guess it

Poke fast:
When working in a SAB we want to make movements in a swift and controlled manner to avoid stirring up too much contaminated air. Don't leave the lids off the agar and LC too long or you will welcome contaminates in.

Poke smart:
When you do your poke try to select mycelium from areas with most desired growth. Also don't go poking around in contaminated agar dishes now.. c'mon you know better than that!


--------------------
:amanita2: Substrate surface conditions / Monotub prep and care :sporedrop:

Look around you... Everything you see exists inside the mind.
Consciousness, the awareness that is experiencing this mind,
is peering in from outside the universe.

Our individual experiences are all part of the universe's experience of itself

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InvisibleComebackKid
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Re: The Biopsy Method: A different way of doing LC. [Re: Prietenul]
    #26445690 - 01/22/20 06:23 AM (4 years, 2 months ago)

One sec


--------------------
:amanita2: Substrate surface conditions / Monotub prep and care :sporedrop:

Look around you... Everything you see exists inside the mind.
Consciousness, the awareness that is experiencing this mind,
is peering in from outside the universe.

Our individual experiences are all part of the universe's experience of itself

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OfflineThe Mycologist
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Re: The Biopsy Method: A different way of doing LC. [Re: ComebackKid]
    #26445691 - 01/22/20 06:24 AM (4 years, 2 months ago)

Haha yep :whathesaid:


I was just quoting the op because he had me laughing with that.


--------------------
"That you are here—that life exists, and identity;
That the powerful play goes on, and you will contribute a verse.”
― Walt Whitman, Leaves of Grass
:acidfire::tmckenna:

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OfflinePrietenul
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Re: The Biopsy Method: A different way of doing LC. [Re: The Mycologist]
    #26445705 - 01/22/20 06:40 AM (4 years, 2 months ago)

Ah I see :lol: That is true

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InvisibleComebackKid
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Re: The Biopsy Method: A different way of doing LC. [Re: Prietenul]
    #26445719 - 01/22/20 07:02 AM (4 years, 2 months ago)

Quote:

Prietenul said:
Quote:

ComebackKid said:
There are no nutrients in the syringe. Just water.




If I put nutes (LC broth) inside it, there are nutes in the syringe. :lol: You read my post, didn't you?

I guess you must have just skimmed over my post. Not a problem. Would be nice if you read it again and give me your opinion.

Edit: Ah it's possible that I wrote too clumsy. I meant, that I put broth in the syringe and the pint jar, before PCing. My English can be improved...and I should go to sleep :lol:





I did read your post and I thought that you were confused about the method. I didnt realize you were proposing a new method. Your english is fine, my bad.

One concern I have with your method is that the nutrient water in the syringe will boil out of the syringe in the PC. You might want to give them a good wipe with alcohol after the PC and before you do any more work with them. (+ contam vector)
Maybe leave the caps off so you dont get nutrients on the inside of the plastic needle cap? (+ contam vector)

I suggest the jar of water have no nutrients at all. You already have nutrients inside the syringe that should be pleanty.
After your poke just open the lid of the jar and draw enough water to get the piece of agar inside.

Also note that the jar has a much better seal than syringe. (+ contam vector)


Not to say your method wont work though. Good luck and let us know how it goes :thumbup:

Edit:
Now that I'm thinking of it more...
If I were to try it this way I might just sterilize an empty syringe. Poke. And draw nutrient water from the jar. But you'd still have to clean the tip after.
Maybe a second jar with no nutrients to top up? Or a wipe with alcohol (+contam vector +contam vector)

Josex method keeps looking better and better


--------------------
:amanita2: Substrate surface conditions / Monotub prep and care :sporedrop:

Look around you... Everything you see exists inside the mind.
Consciousness, the awareness that is experiencing this mind,
is peering in from outside the universe.

Our individual experiences are all part of the universe's experience of itself

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OfflinePrietenul
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Re: The Biopsy Method: A different way of doing LC. [Re: ComebackKid]
    #26445747 - 01/22/20 07:23 AM (4 years, 2 months ago)

Quote:

ComebackKid said:
Quote:

Prietenul said:
Quote:

ComebackKid said:
There are no nutrients in the syringe. Just water.




If I put nutes (LC broth) inside it, there are nutes in the syringe. :lol: You read my post, didn't you?

I guess you must have just skimmed over my post. Not a problem. Would be nice if you read it again and give me your opinion.

Edit: Ah it's possible that I wrote too clumsy. I meant, that I put broth in the syringe and the pint jar, before PCing. My English can be improved...and I should go to sleep :lol:





I did read your post and I thought that you were confused about the method. I didnt realize you were proposing a new method. Your english is fine, my bad.





No problem at all. :smile:

Quote:


One concern I have with your method is that the nutrient water in the syringe will boil out of the syringe in the PC. You might want to give them a good wipe with alcohol after the PC and before you do any more work with them. (+ contam vector)
Maybe leave the caps off so you dont get nutrients on the inside of the plastic needle cap? (+ contam vector)





Yeah, I was expecting a little boil over. The idea to leave the cap off is good, though. I was thinking, that I have to pull less broth into the syringe if it is over half full after PCing, which would reduce the contam vector. I would not have to tilt the jar and so on...

Quote:


I suggest the jar of water have no nutrients at all. You already have nutrients inside the syringe that should be pleanty.
After your poke just open the lid of the jar and draw enough water to get the piece of agar inside.





This is even better. You're right. No need for more nutrients.

Quote:


Also note that the jar has a much better seal than syringe. (+ contam vector)





Sure thing. I planned to poke, pull water, flame sterilize the needle,  then cap it and put the syringe in a ziplock-bag. Everything inside the SAB of course. This should keep the syringe sterile until used, right?

Quote:


Not to say your method wont work though. Good luck and let us know how it goes :thumbup:





Thanks ComebackKid, I will surely report the results.

Quote:


Edit:
Now that I'm thinking of it more...
If I were to try it this way I might just sterilize an empty syringe. Poke. And draw nutrient water from the jar. But you'd still have to clean the tip after.




The downside of this is, that I'd again have to pull the 60 ml into the syringe (4 times), and thus either use many jars with nutrients or less big ones, which is a hassle to pull from cleanly. I would clean the tip in any case by just flame sterilizing it after everything is done. Then caping and putting in ziplock-bag.

Quote:


Maybe a second jar with no nutrients to top up? Or a wipe with alcohol (+contam vector +contam vector)

Josex method keeps looking better and better




I agree, that this method seems strange... I'll have to think a little bit more and make a plan. Your inputs have been of great value anyway. Thank you very much. ...and expect my results to appear soon. :smile:

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Offlinetrubblesome
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Re: The Biopsy Method: A different way of doing LC. [Re: Prietenul]
    #26446001 - 01/22/20 10:01 AM (4 years, 2 months ago)

dont use fire in the SAB!


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

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OfflinePrietenul
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Last seen: 3 years, 6 months
Re: The Biopsy Method: A different way of doing LC. [Re: trubblesome]
    #26446075 - 01/22/20 10:36 AM (4 years, 2 months ago)

Quote:

trubblesome said:
dont use fire in the SAB!




Of course not :smile: It would be too small anyway. I have the burner next to it standing on a sink. I usually flame until it glows red and immediately put it into the SAB. That's a distance of maybe 30cm...

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Offlinetrubblesome
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Re: The Biopsy Method: A different way of doing LC. [Re: Prietenul]
    #26446079 - 01/22/20 10:38 AM (4 years, 2 months ago)

okay phew!!


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

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