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InvisibleKingKobra
Snake


Registered: 10/02/20
Posts: 4
Skip the LC: Inoculate BRF directly with Josex poke and remove a contam vector * 1
    #27078116 - 12/08/20 02:50 AM (3 years, 2 months ago)

Recently, I was having trouble getting started with a grow, and I wanted to turn 2 successful BRF jars into more. I started a thread about it, and the consensus seemed to be: clone a pin to agar, and then make LC using the Josex poke method.

I did make LC as advised in the original thread, but—just to try it—I also inocc'd 5 jars directly by poking the agar with the syringe needle, and squirting into the BRF jars as if I was doing PF tek. By the time the LC was ready, there was already visible growth in the jars in all spots I inoculated, except for 1. At 4 holes per jar, and 5 jars, that is a 95% success rate, with no contams to-date (it's been a few weeks). Since the method is a bit counter intuitive but seemed to work, I figured I would share.

My exact steps:
  • Clone a pin to agar and wait for happy, healthy mycelium
  • Fill some syringes with sterile water using this 10-year old method
  • Flame sterilize the needle (between jars)
  • In a SAB: Poke the mycelium on agar with the sterile water syringes as per Josex poke
  • In a SAB: Insert the syringe into the BRF jar like in PF tek, and squirt out a generous amount of water. I used one 10mL syringe of water for 2 jars with 4 holes each, so that's roughly 2.5cc per squirt. But it doesn't really matter as long as you're generous
  • ...:mushroom2:


Pros of this method:
  • No need for a PC to sterilize the liquid culture, or anything else (this came in handy since my PC was out of commission for a while)
  • One less vector of contamination – LC can be scary
  • Don't have to take out and set up the SAB twice for inoculating LC and inoculating jars – only need to set it up once to poke the agar and inoculate the jars in one go
  • Growth has been much quicker than with spores, but I don't have enough experience to compare directly with LC here. I went from inoculation to full colonization in 2.5 weeks
  • Seems to be fairly high success rate and fairly low risk (despite small sample size)
  • Overall just really easy


I don't feel like I've had enough mushroom-growing experience to evaluate the cons, but hopefully this bit of tek is helpful for anyone with agar who needs to knock up BRF cakes! Would love for someone to replicate and confirm for others that this works.


Edited by KingKobra (12/08/20 02:52 AM)


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InvisibleJosex
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Registered: 11/13/15
Posts: 8,995
Loc: Flag
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Re: Skip the LC: Inoculate BRF directly with Josex poke and remove a contam vector [Re: KingKobra]
    #27078227 - 12/08/20 05:55 AM (3 years, 2 months ago)

Can't see no cons myself, you're getting faster growth from this than with spore solution with the added benefit of inoculating with clean tissue sample, because we know spore solution is always a bit of a gamble.

I'm glad that you are excited about it and that it's working for you without a PC. When results come, you could flesh this out into a tek. Pretty sure more people without a PC can find this interesting. I would do LC myself even wihtout a PC though, it's perfectly doable.


Edited by Josex (12/08/20 06:12 AM)


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InvisibleKingKobra
Snake


Registered: 10/02/20
Posts: 4
Re: Skip the LC: Inoculate BRF directly with Josex poke and remove a contam vector [Re: Josex]
    #27079862 - 12/09/20 03:23 AM (3 years, 2 months ago)

Could see writing up a quick tek with some data once things shake out completely.

Any particular reason you'd still stick to LC?


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InvisibleShroombitz
Not entitled to a title
I'm a teapot User Gallery

Registered: 11/19/20
Posts: 176
Loc: Europe
Re: Skip the LC: Inoculate BRF directly with Josex poke and remove a contam vector [Re: KingKobra]
    #27080458 - 12/09/20 12:48 PM (3 years, 2 months ago)

Quote:

KingKobra said:
Could see writing up a quick tek with some data once things shake out completely.

Any particular reason you'd still stick to LC?




Only works for PF-Tek and/or is a PITA when doing more than a couple of jars?


--------------------
Ask not what your shrooms can do for you, ask what you can do for your shrooms.


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