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Offlinespock1
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fruit biopsy - iodine? *DELETED*
    #7604221 - 11/06/07 07:27 PM (16 years, 4 months ago)

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InvisibleThePyschonaut52
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Re: fruit biopsy - iodine? [Re: spock1]
    #7604239 - 11/06/07 07:32 PM (16 years, 4 months ago)

DON'T USE IODINE.

It kills the tissue...from my experience


--------------------
"In god we trust..."


-I guess we're screwed.

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Invisiblemycocurious
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Re: fruit biopsy - iodine? [Re: ThePyschonaut52]
    #7604248 - 11/06/07 07:35 PM (16 years, 4 months ago)

I use iodine rubs over the tissue before biopsies for culture isolates. It kills off most contaminates and doesn't seem to stress the tissue out as much as H2O2 does as the mycelium recovers in agar sooner with these than with H2O2 dipped biopsies...


--------------------
:justdontknow: Don't mistake my tone for a "matter-of-fact" attitude.  I'm just presenting what I believe to be correct, until I'm corrected...

- How Myco-Curious Prepares Coir & Compost Substrates
- How Myco-Curious Builds A Bulk Humidifier
- How Myco-Curious Builds An Automated Greenhouse
------------------------------------
figgusfiddus said:
Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.

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Offlinespock1
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Re: fruit biopsy - iodine? *DELETED* [Re: ThePyschonaut52]
    #7604257 - 11/06/07 07:36 PM (16 years, 4 months ago)

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InvisibleThePyschonaut52
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Re: fruit biopsy - iodine? [Re: mycocurious]
    #7604260 - 11/06/07 07:37 PM (16 years, 4 months ago)

Hmph, probably just me. I used a 100% iodine solution though...


--------------------
"In god we trust..."


-I guess we're screwed.

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OfflineRogerRabbitM
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Re: fruit biopsy - iodine? [Re: spock1]
    #7604270 - 11/06/07 07:39 PM (16 years, 4 months ago)

You shouldn't need to do anything but clean the surface if you're using that tek. In fact, skip all the cleaners and just peel off a strip of mycelium like a banana peel, and then stick the needle into the freshly exposed flesh. Iodine is fine for cleaning the outside of the tissue, but use it at no more than ten percent.
RR


--------------------
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semper in excretia sumus solim profundum variat

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Offlinehightimesreader
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Re: fruit biopsy - iodine? [Re: RogerRabbit]
    #7604369 - 11/06/07 08:06 PM (16 years, 4 months ago)

So, you iodine the surface of the mushroom, then you peel off that layer, and scalpel out a peice and put it on a hardened agar plate? For an LC, you'd scoop out fresh tissue and fill the bore of the needle a section of myc. right? How you get it out?

HTR


--------------------
I'm hunting for The Following ethnos.
For experiments, hunting finds and any other contributions, check out My journal.
HTR

A new leaf turned over.. I'm too old for this shit.
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Invisiblemycocurious
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Re: fruit biopsy - iodine? [Re: hightimesreader]
    #7604431 - 11/06/07 08:17 PM (16 years, 4 months ago)

That's the method I follow although if I was doing a liquid culture I would take a needle and flame sterilize it and while red hot, draw in a few cc's of air through the super-heated air of the flame. (sterilized air)

I would then plunge the needle into the exposed tissue and twist while pulling up on the plunger ever so slightly. That will cut a plug into the syringe. Then just place it into the LC and depress the plunger to "shoot" it in to the liquid medium.

Repeat by beginning at the flame sterilization procedure again...


--------------------
:justdontknow: Don't mistake my tone for a "matter-of-fact" attitude.  I'm just presenting what I believe to be correct, until I'm corrected...

- How Myco-Curious Prepares Coir & Compost Substrates
- How Myco-Curious Builds A Bulk Humidifier
- How Myco-Curious Builds An Automated Greenhouse
------------------------------------
figgusfiddus said:
Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.

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Invisiblejeetered
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Re: fruit biopsy - iodine? [Re: ThePyschonaut52]
    #7604600 - 11/06/07 08:46 PM (16 years, 4 months ago)

Quote:

ThePyschonaut52 said:
DON'T USE IODINE.

It kills the tissue...from my experience




I've had lots of new success using providone-iodine (very hard to find pure iodine)

but, u can take providone, mix it with 3 times the amount of h2o2, and it will precipitate iodine crystals, that u can dilute with dh2o, and have pure iodine. LOL.


k,

my 2 cents.

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Invisibleblood4blood
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Registered: 04/25/07
Posts: 6,029
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Re: fruit biopsy - iodine? [Re: jeetered]
    #7604676 - 11/06/07 09:00 PM (16 years, 4 months ago)

what is the best gauge syringe to use for this? can i get away with an 18 gauge? because thats all i have

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Invisiblemycocurious
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Re: fruit biopsy - iodine? [Re: blood4blood]
    #7604767 - 11/06/07 09:16 PM (16 years, 4 months ago)

that's fine, just take several plugs from a very (relatively) close area on the fruit and flame sterilize between every single extraction.

because a single fruit can still contains multiple sub-strain genetics you want your biopsies to be very densely packed.


--------------------
:justdontknow: Don't mistake my tone for a "matter-of-fact" attitude.  I'm just presenting what I believe to be correct, until I'm corrected...

- How Myco-Curious Prepares Coir & Compost Substrates
- How Myco-Curious Builds A Bulk Humidifier
- How Myco-Curious Builds An Automated Greenhouse
------------------------------------
figgusfiddus said:
Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.

Extras: Filter Print Post Top
Offlinespock1
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Re: fruit biopsy - iodine? *DELETED* [Re: mycocurious]
    #7604815 - 11/06/07 09:26 PM (16 years, 4 months ago)

Post deleted by spock1

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Invisiblemycocurious
Mike O. Kuerias
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Registered: 02/09/07
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Re: fruit biopsy - iodine? [Re: spock1]
    #7604835 - 11/06/07 09:33 PM (16 years, 4 months ago)

well, there is strength in numbers, make sure to have at least several in case one or more of them do not grow out.


--------------------
:justdontknow: Don't mistake my tone for a "matter-of-fact" attitude.  I'm just presenting what I believe to be correct, until I'm corrected...

- How Myco-Curious Prepares Coir & Compost Substrates
- How Myco-Curious Builds A Bulk Humidifier
- How Myco-Curious Builds An Automated Greenhouse
------------------------------------
figgusfiddus said:
Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.

Extras: Filter Print Post Top
Offlinespock1
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Registered: 04/14/07
Posts: 589
Last seen: 15 years, 9 months
Re: fruit biopsy - iodine? *DELETED* [Re: mycocurious]
    #7609082 - 11/07/07 10:00 PM (16 years, 4 months ago)

Post deleted by spock1

Reason for deletion: .


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Invisibleblood4blood
Calmer Than You Are


Registered: 04/25/07
Posts: 6,029
Loc: The Valley
Re: fruit biopsy - iodine? [Re: spock1]
    #7609108 - 11/07/07 10:07 PM (16 years, 4 months ago)

Quote:

spock1 said:
I filed down a few needles to 90 degrees, and tested it out on a fruit stem. They take a little piece like 1 mm (about 1/32") long. Is that really enough to start a 250ml (about a cup) or larger LC?

I got a piece from the cap that was 2 mm (about 1/16") long. Can I clone tissue from the cap?




i was wondering this also if you can clone from the cap or just the stem.. in RR's video he shows you cloning from both but those are wild edibles i dont know if cubensis would be any different.



SPOCK1- what did you end up wiping down the mushroom with before you took your core?

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Invisiblemycocurious
Mike O. Kuerias
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Registered: 02/09/07
Posts: 1,265
Re: fruit biopsy - iodine? [Re: spock1]
    #7609111 - 11/07/07 10:08 PM (16 years, 4 months ago)

yes and yes. just take several plugs from a very close area to ensure both similar genetics between the samples (multiple genetics can exist in a single fruit so cloning this way is kind of tricky) and to ensure that you have at least one biopsy that proves viable and starts growing out in the liquid medium. two or three plugs are all you need per 250ml LC to inoculate it...one would be fine, but the risk of it not growing out is higher.


--------------------
:justdontknow: Don't mistake my tone for a "matter-of-fact" attitude.  I'm just presenting what I believe to be correct, until I'm corrected...

- How Myco-Curious Prepares Coir & Compost Substrates
- How Myco-Curious Builds A Bulk Humidifier
- How Myco-Curious Builds An Automated Greenhouse
------------------------------------
figgusfiddus said:
Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.

Extras: Filter Print Post Top
Invisibleblood4blood
Calmer Than You Are


Registered: 04/25/07
Posts: 6,029
Loc: The Valley
Re: fruit biopsy - iodine? [Re: mycocurious]
    #7609119 - 11/07/07 10:09 PM (16 years, 4 months ago)

is it best to rip the cap in half and take a core from the tissue there or go right up through the gills?

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Invisiblemycocurious
Mike O. Kuerias
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Registered: 02/09/07
Posts: 1,265
Re: fruit biopsy - iodine? [Re: blood4blood]
    #7609138 - 11/07/07 10:15 PM (16 years, 4 months ago)

that has already been discussed in the beginning of the thread... after washing the exterior of the fruit with iodine, peel back the skin of the cap and take the biopsy from that exposed, sterile flesh.


--------------------
:justdontknow: Don't mistake my tone for a "matter-of-fact" attitude.  I'm just presenting what I believe to be correct, until I'm corrected...

- How Myco-Curious Prepares Coir & Compost Substrates
- How Myco-Curious Builds A Bulk Humidifier
- How Myco-Curious Builds An Automated Greenhouse
------------------------------------
figgusfiddus said:
Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.

Extras: Filter Print Post Top
Invisibleblood4blood
Calmer Than You Are


Registered: 04/25/07
Posts: 6,029
Loc: The Valley
Re: fruit biopsy - iodine? [Re: mycocurious]
    #7609152 - 11/07/07 10:19 PM (16 years, 4 months ago)

ah sorry myco i thought that was just in reference to taking a core from the stem.. i asked a question earlier that didnt get answered which was syringe size all i have is an 18 gauge will that work or should i go for something bigger???

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Invisiblemycocurious
Mike O. Kuerias
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Registered: 02/09/07
Posts: 1,265
Re: fruit biopsy - iodine? [Re: blood4blood]
    #7609160 - 11/07/07 10:24 PM (16 years, 4 months ago)

yea, it's fine.. a couple plugs extracted as described above works fine. I'd have to double check the archives but I vaguely recall there being less chance of multiple genetics when samples are taken from the cap rather than the stem...

easier to discern in an agar plate than in a liquid culture though as you can observe the growth in a more 2-dimensional view...


--------------------
:justdontknow: Don't mistake my tone for a "matter-of-fact" attitude.  I'm just presenting what I believe to be correct, until I'm corrected...

- How Myco-Curious Prepares Coir & Compost Substrates
- How Myco-Curious Builds A Bulk Humidifier
- How Myco-Curious Builds An Automated Greenhouse
------------------------------------
figgusfiddus said:
Keep in mind that inoculating or whatever in front of a flow hood won't help your bad substrate, your bad inoculant, your bad sterile procedure, etc. etc. etc. It's not a +3 flowhood of magic, it's just a tool.

Extras: Filter Print Post Top
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