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Invisiblesporealchemy
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Registered: 04/10/11
Posts: 157
Loc: mf mars my guy
Re: Easiest Liquid Culture for the incredibly lazy [Re: elasticaltiger]
    #25120376 - 04/07/18 11:49 AM (4 months, 6 days ago)

Quote:

elasticaltiger said:
Whenever a transfer piece bounces around on the agar I can see growth at every spot where the wedge hit the agar even though there was no visible mycelium on the spots.

I've been contamplating for a while meow whether it would be possible to clone a fruit just by rubbing a sterile swab on the inside. Never had the time to try it out though.




I've noticed this happening even tho i hate when a pin rolls around in a dish, but there are often multiple clean colonies.

And I'd bet that sterile swab would totally work and would like to try this also wow


--------------------
"Once your spawn jars are fully colonized, you can scratch your butt while you inoculate the coir if you want."
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Invisibleplurfekt
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Re: Easiest Liquid Culture for the incredibly lazy [Re: Pastywhyte]
    #25120726 - 04/07/18 02:41 PM (4 months, 6 days ago)

Quote:

Pastywhyte said:
Don't underestimate a cells ability to divide. I cloned a fruit once just by dragging a blade through the tissue, did not remove a visible amount of tissue. Then dragged that blade through agar and had growth.




I'm tryin it.

That's dope :thumbup:

BTW - thank you very much man; I'm now working with agar and have my first clean pasty plates colonizing; immediately went out and bought more mini rounds lol. :hug:


Edited by plurfekt (04/07/18 02:44 PM)


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OfflineDr. Freeman
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Re: Easiest Liquid Culture for the incredibly lazy [Re: plurfekt]
    #25136749 - 04/13/18 09:54 PM (3 months, 30 days ago)

Post edit:
I added the Grocery Store Agar Tek to my thread.
I think it would be a really good complement for any LC.


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OfflineGrimsweeper
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Re: Easiest Liquid Culture for the incredibly lazy [Re: Dr. Freeman]
    #25142099 - 04/15/18 11:04 PM (3 months, 28 days ago)

Doesn't appear that we can buy pre-filled sterile dextrose IV bags in the US. Just wondering your thoughts on filling an empty bag like this..

https://www.amazon.com/KangarooGravity-Feeding-Large-Bore-1000mL/dp/B00T5AFVDW/ref=sr_1_fkmr2_1?ie=UTF8&qid=1523846209&sr=8-1-fkmr2&keywords=iv+bag+autoclave

with a homemade LC then PC the whole thing.
Only thing I'm uncertain of is if there's an inoculation port on this bag or if you'd need something like this as well..

https://www.amazon.com/Baxter-IV-Set-Extra-Port/dp/B003VP2MR0/ref=pd_bxgy_328_img_2?_encoding=UTF8&pd_rd_i=B003VP2MR0&pd_rd_r=XS9SB7TVHHXTGPK64WFK&pd_rd_w=ymj9c&pd_rd_wg=s1jjW&psc=1&refRID=XS9SB7TVHHXTGPK64WFK


--------------------
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OfflineDr. Freeman
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Re: Easiest Liquid Culture for the incredibly lazy [Re: Grimsweeper]
    #25143345 - 04/16/18 03:08 PM (3 months, 28 days ago)

I'm not sure the material of the bag would withstand the temperature of a PC.
And even if it did, a completely closed bag with boiling liquid doesn't seem like a good idea.
Probably the reason they use radiation to sterilize.

Post edit
I think making an IV bag defeats the purpose of buying pre-made.
Go with jars instead.


Edited by Dr. Freeman (04/16/18 03:11 PM)


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OfflineConntrap
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Re: Easiest Liquid Culture for the incredibly lazy [Re: Dr. Freeman]
    #25306959 - 07/03/18 08:52 PM (1 month, 11 days ago)

Hey Doc, (people)

I've read your post a couple hours ago and it inspired me to give it a try. Due to the different Dextrose I could buy i had to change the tek a little bit.

Quote:

Dr. Freeman said:
On a sidenote, I prepare several syringes with a tissue sample and 2.5% dextrose (1 part 5% dextrose and 1 part sterile water).
I only inject back the syringes that show grow after a few days.
This is about 30%.

I find it better to waste a few syringe than a whole LC.
Never tried this from a Spore Syringe of my own making, success rate wouldn't be that great.





After reading that I thought, why not? I prepared two sterile containers, one with a multi spore syringe and the other with the 5% dextrose (not diluted in more water). Always inside a SAB. Afterwards I whitdrawed 3ml of dextrose from one container and then 2ml of spore solution.
What do you think ? Where should I store the syringes ? Should micelium grow inside each syringe if no contamination appears ? If yes, I understand that i could inyect that colonized liquid (Liquid culture already?)into sterilized grain ?



This was originally a PM but I decided to share it with all of you reading.-


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OfflineDr. Freeman
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Re: Easiest Liquid Culture for the incredibly lazy [Re: Conntrap]
    #25307577 - 07/04/18 06:34 AM (1 month, 11 days ago)

Quote:

Conntrap said:





Dextrosa al 5% en agua.
It certainly seems easier to buy in Hispanic countries.
:fistbump:
2 ml of spores in 3 ml dextrose is an incredibly large amount of spores.
1 drop would have sufficed.
Spores should germinate in those syringes whether you have contaminants or not.
I would try a few drops on agar. If agar is clean, make a larger LC.
You can use the whole IV bag or you can make at least a 250cc LC in a pint jar.
Make 25 syringes out of that.
Use to inoculate grain.

I recently tried spores > LC syringe > agar. The LC looked good and healthy. Tried it on agar, green stuff grew :thumbdown: If I had tried grains directly I would have wasted a lot of resources and TIME.

How is your agar game?
I have found it is a vital tool in the fight against contaminants.
:paranoid: You can never be too careful in your fight against green fungus. :paranoid:

Post edit:

What works for me: Spores > Agar > LC > Grains
The "Spores, agar, LC" might take a little time, but with one drop of spores using this method; you could end up inoculating 100 pounds of grains.


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OfflineConntrap
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Re: Easiest Liquid Culture for the incredibly lazy [Re: Dr. Freeman]
    #25344902 - 07/24/18 07:25 PM (22 days, 40 minutes ago)

Quote:

Dr. Freeman said:
Quote:

Conntrap said:





Dextrosa al 5% en agua.
It certainly seems easier to buy in Hispanic countries.
:fistbump:
2 ml of spores in 3 ml dextrose is an incredibly large amount of spores.
1 drop would have sufficed.
Spores should germinate in those syringes whether you have contaminants or not.
I would try a few drops on agar. If agar is clean, make a larger LC.
You can use the whole IV bag or you can make at least a 250cc LC in a pint jar.
Make 25 syringes out of that.
Use to inoculate grain.

I recently tried spores > LC syringe > agar. The LC looked good and healthy. Tried it on agar, green stuff grew :thumbdown: If I had tried grains directly I would have wasted a lot of resources and TIME.

How is your agar game?
I have found it is a vital tool in the fight against contaminants.
:paranoid: You can never be too careful in your fight against green fungus. :paranoid:

Post edit:

What works for me: Spores > Agar > LC > Grains
The "Spores, agar, LC" might take a little time, but with one drop of spores using this method; you could end up inoculating 100 pounds of grains.





Well, actually only 2 syringes out of 15 have some mycelium growing, other 2 are contaminated (green spot) and the rest look exactly the same as the day I prepared them. That really surprised me since i used two prints for the 15 syringes, diluted. I think it's more than enough and I don't think there's a max amount of spores per mililiter of liquid.
I have some agar to pour, will be testing it soon. I'm not that good with the inoculation loop, I have to practice and watch some videos to learn


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OfflineDr. Freeman
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Re: Easiest Liquid Culture for the incredibly lazy [Re: Conntrap]
    #25345872 - 07/25/18 11:50 AM (21 days, 8 hours ago)

It really isn't surprising.
That high failure rate might be because there are contaminants in the syringe and/or you introduce em while manipulating.
The problem is most probably the spores.

I do Spores > Agar > Poke-biopsy to LC in syringes.
Works considerably better than spores > LC.


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