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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Violet]
    #24809791 -

You critique his time frame when thats missing the point stated in the title: achieving clean bacteria free spawn.


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Re: The Biopsy Method: No Mercy for Bacteria. [Re: JHOVA]
    #24810561 -

I missed no point, I'm fully aware of that.  Seems you didn't understand my reason for saying that but it's okay, you might understand from my response to him.


Josex,
pretty much any myc torn from their culture is somewhat harmed. That's not entirely a bad thing of course, we do it all the time on purpose.  But making "homeless" harmed myc establish and recover is much slower than transferring something with myc mostly safely protected inside, like a grain or wedge. Your extra time you mentioned is the myc recovering, then establishing itself in new food, before it can begin to grow out. When the effect of leaving bacteria behind is the same, why use the method that makes the myc take extra time to recover, especially when it spreads (at least more than a wedge etc) the contamination you're trying to run away from?

I figure you're likely fairly right about the water spreading bacteria on PF sub, but nonetheless this will spread it more than a wedge, and when a method takes extra (unnecessary?) time to recover And involves a lot more cellular expansion, I guess I wanna know... why do it?

The "cross-contam" comment was kindof a throwback to the joke I made in the other thread.  The "biopsy" poke essentially works on the dynamics of cross-contamination, I wasn't saying you're cross-contamming, just referring to the biopsy as "cross-contam on purpose", lol.

I'm not really critiquing.  I'm mostly trying to understand why this is preferable, as well as why it's being written up in this particular way. Honestly it seems like learning a new clever trick but downgrading other aspects of the process to justify using it. You remember me commenting that the biopsy poke was clever and I may have some use for it in my culturing (although I did take note of your concern, that just loose cells may not take hold well on grain petris, an insight that I bet is correct and I'm glad you mentioned, I just forgot to respond).  Just trying to understand the rationale for preferring to use the biopsy to escape contams instead of method that favors the myc even more.


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Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
• Violet's Teks and Posts •

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Violet]
    #24810566 -

FWIW this is coming from someone with effectively Zero problems with contams.


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Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
• Violet's Teks and Posts •

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Violet]
    #24810640 -

Surprisingly I had taken a poked piece to a quart of grain on 11/13. Just noticed some growth yesterday.


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If I ever give out misinformation please inform me so I can have the correct information. :cheers:

Tmethyl said:
Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy.

Caps McGee said:
:thumbsup:
Fun part is figuring out what works best for you

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: van hatton] * 1
    #24810733 -

Violet, you're probably right about noccing with a wedge, the mushrooms gods have told me that while I was playing Skyrim.

But... you gotta trust me when I tell you that the last thing you'd feel like doing if you had had such disgusting swabbed plates on your hands as I had is taking a damn wedge from it. Maybe a wedge can work with a less severe contamination or maybe even with heavy who knows, although I doubt it. Will try for sure when the occasion presents itself and will report back.

I don't care about celular expansion and all the theory tbh. All I really know is that the grows where I used wedges to inoculate paled in comparison with the ones from LC using the poke. You could say it is just coincidence but I know very well it isn't. I could swear there's something about the fucking poke but don't know exactly what it is.

Get a nice clean agar culture with your trusty scalpel and poke the hell out of it to make as much LC as you need. That's all I care about.

You make it sound almost as if the celular division from a single poke is tantamount to doing a protracted g2g expansion... it isn't that big of a deal.
The colony will recover just fine from a poke.
What about people investing months of their time on getting an isolate, cos that's celular division right there. Nonetheless, good isolates will perform very well.


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LC's are easy
Go big with No-pours
Clean yo shit
Has u got Magic?

PHENO HUNTERS

Edited by Josex (12/11/17 03:19 PM)

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: van hatton]
    #24810754 -

van hatton said:
Surprisingly I had taken a poked piece to a quart of grain on 11/13. Just noticed some growth yesterday.


:mostinteresting:
Keep me posted, that sounds like it's gonna take too long to colonize though.


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LC's are easy
Go big with No-pours
Clean yo shit
Has u got Magic?

PHENO HUNTERS

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Josex]
    #24811199 -

I understand the march towards senescence isn't a big deal for most growers, especially when Cubensis can do so well from multi-spore thanks to bulk subs. I mostly mention the cell expansion as an aside, I don't blame you for not caring much, you're right not to.  I personally consider it because I like holding on to cultures with just a few inches worth of radial expansion.  And you know me, I just like bringing up any and all relevant facts sometimes. Especially since this isn't per se a Cubensis tek... Losing edible cultures to senescence is a much bigger deal.


Imagined the feeling of purposefully sticking my scalpel right into a bacterial blotch.  Gross. You've got me with you on your page with that now.  I do hate contams.  I do well to never have to mess with bacteria, and when I did I used gentamicin sulfate to fair effect, but indeed there's no need for antibiotic agar when something like my grain petri or a brf/verm dish does the job.

That aside, if I were using the substrate run-away trick I would want to know exactly where I was placing the contam and also give myc the faster jump-off. Gross as it might feel, I'd still use the wedge. Mycelium need to make itself a 'home' to colonize from, and bacteria doesn't, which would give bacteria an advantage when you inoc mycelium in a way to where it has to find that new 'home' before it can compete. Myc have an advantage on the new media, which is what this tek is counting on, IMO perhaps more than it should have to.

But of course that's just describing what I would do or suggest and why.
I have no reason to think this wouldn't work.
Although I do wonder if sometimes those very few 'homeless' myc cells might not get a foothold on sub shared with bacteria, similar to your concern with biopsy for my grain petris with the added concern of bacterial contams.  But seeing as everyone's LCs have taken off nicely already, I would be surprised if people report many cases of no growth.

And as I said before I haven't had to clean away from a contam in years.  Ziploc containers and sterile airflow are such a luxury...


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
• Violet's Teks and Posts •

Edited by Violet (11/28/17 10:22 AM)

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Violet]
    #24811794 -

Quote:
Although I do wonder if sometimes those very few 'homeless' myc cells might not get a foothold on sub shared with bacteria, similar to your concern with biopsy for my grain petris with the added concern of bacterial contams.


Sometimes the biopsy won't get a foothold on pf-tek sub, it's not frequent but it happens, regardless of the tissue squirted being clean or not.
On the other hand, I'm pretty sure the biopsy will grow 100% of the times in a liquid medium, it has not failed me a single time yet. The tissue does not need any foothold in the broth since it's floating in a nutrient liquid medium.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
Has u got Magic?

PHENO HUNTERS

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Josex]
    #24811844 -

Yes exactly.  I have a new set of multi-spore germ plates, thinking about using a blunt wide gauge to see how it takes to grain petri for part of this run, if I do I'll let you know


--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
• Violet's Teks and Posts •

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Violet]
    #24813452 -

Josex, what's the hole in the lid for? If you're using the screw top ziplocs, you can unscrew the lid a little. If you're using mason jars they can stand up to the PC pressure.

It seems like if you're just growing clean myc out you don't need it. Maybe you need it for fruiting in-vitro?

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: senseit]
    #24813459 -

senseit said:
you need it for fruiting in-vitro


Exactly. If it was agar there wouldn't be a problem with no hole in the jar but it is a hot sub that can pin prolifically.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
Has u got Magic?

PHENO HUNTERS

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Josex]
    #24813676 -

Very cool write up :thumbup:


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"A single lifetime in this density can accomplish a refinement of Will that might take far longer in a realm where the truth is always visible" - Zii, Confederation of Planets, GFL Station.

"You will then have national and global contact around 2026." - Pleiadian Prophecy 2020

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Josex]
    #24912080 -

This needs a bump. Clean write up brother and a good new update, Thanks for sharing.
I wish I dug on to this earlier before wasting 50  agar dishes & time trying to clean a fuckin dirty spore print from bacteria.

Pucks are essential to this, since Ive been using poke transfering tissue from agar to agar with no success. Bacteria really loves the smooth agar surface.

I may restart from prints directly to pucks.
Just lacks verm as of the moment. Have plenty of coir though.
-Can verm be substitute to coir for the pf recipe?

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: pacmanbreed] * 1
    #24912197 -

I'm glad it helped man. :thumbup:

Yes, you can do coir+brf pucks but you need to adjust the ratios, way less brf for starters. You can bucket tek the coir separately and then add the brf, 1:5 ratio (brf:coir) is perfect.

Coir pucks are not nearly as effective as PF pucks for getting rid of bacteria, though. But that doesn't mean they aren't effective, just not as much. They're also good if for whatever reason you're feeling overwhelmed by your agar workload at any given moment because they grow slowly, also good to take some clones when they pin out.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
Has u got Magic?

PHENO HUNTERS

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Josex] * 1
    #24912959 -

Nice tek and very nice write up Josex :bow2:
Missed this one earlyier

I am going to try a modified version of your technique; I had no usable jars, so I filled some glass petris with BRF mix. They are in the oven as we speak for 65 minutes at 260F/125C

I am going to try to clean up cultures that are proven to be extremely fucking hard to clean. So if this works for those particular cultures it has me smiling!

More later!

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Shroomway] * 1
    #24912965 -

Have you tried sterilizing brf in the oven before with good results? I think they're gonna lose a lot of moisture that way and idk you can sterilize it in the oven. Can't you do steam-sterilization in a pot instead?


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
Has u got Magic?

PHENO HUNTERS

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Josex]
    #24912982 -

Mmm, you might be right... I tried PCing petris, but somehow, (even if double wrapped in foil) water finds it's way into the pretris. This is why I am trying it this way now. It's so much work to do it all over again. Fingers crossed I guess... Thanks though!

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Shroomway]
    #24913004 -

idk man, can't you get small glass jars to do this? Even plastic will work in a pinch.


--------------------
LC's are easy
Go big with No-pours
Clean yo shit
Has u got Magic?

PHENO HUNTERS

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Josex]
    #24913037 -

Yeah, I will for sure get some jars if it looks dark and dried out. Quite some chance that will happen...

Very curious to try this out

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Josex]
    #24913272 -

Josex said:
I'm glad it helped man. :thumbup:

Yes, you can do coir+brf pucks but you need to adjust the ratios, way less brf for starters. You can bucket tek the coir separately and then add the brf, 1:5 ratio (brf:coir) is perfect.

Coir pucks are not nearly as effective as PF pucks for getting rid of bacteria, though. But that doesn't mean they aren't effective, just not as much. They're also good if for whatever reason you're feeling overwhelmed by your agar workload at any given moment because they grow slowly, also good to take some clones when they pin out.


Good to see that we both use the same pp5 plastic container. I love them being small than glads and clear lids.


Thanks for the ratios man. Been wasting 2 months fighting bacteria.
Ill use coir 1:5 ratio and finish it with adjusted ph tea agar. (Hoping to receive my paycheck soon for some verm) :stoned:.
I think your right on coir being slightly less anti bacterial.
I guess its due to the acidic ph of it compared to neutral verm.
- ill wait till this 2nd transfer from spores to pucks will pin for a better clone and isolation.

Heres my plan.
Agar spore streak>pucks>pucks>pins>agar>grain or lc
Or omit streak
Spores direct on puck center> 2nd puck>pins>agar>grain or lc
Hoping to grow & achieve my 1st shoebox using your biopsy methods.

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