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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Re: The Biopsy Method: No Mercy for Bacteria. [Re: pacmanbreed] * 1
    #24913333 -

They aren't exactly the same containers but yours look badass af too. :rockon:

Whatever you do keep us posted man, wish you luck.
If you don't mind me recommending you something, only use the poke when necessary, otherwise transfer normally from puck to puck or from puck to agar, that will speed things up and you won't stretch the culture more than necessary. :thumbup:


--------------------
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]
    #24913380 -

...what do you mean necessary? When do you call it "necessary" to use the poke/biopsy method? I can think of some reasons but are actually curious what your view on this is.. Would you?

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Shroomway]
    #24913392 -

I mean, if you are working with a known clean culture there's no need to take biopsies from them to transfer, it's slow and you have to take into account that such a tiny winy piece of tissue needs to undergo a lot of cellular division to start a colony from scratch, so no need to subject the culture to unnecessary stress.


--------------------
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] * 3
    #24913497 -

Sure will do.
Understood & Noted man. Will limit 21g use upto max of 3 times only.
Shroomway only necesary for bacterial culture or tight corners from contams, No need to use on clean cultures. Thats why its better to limit 3-6 scapel grain size wedge transfers from spores to limit cellular division/age/stress. not all isolate are good fruit bearers. Josex really know his good shit.

Glad ive found your much better write up Considering this.
I cant consider My method as biopsy as of the moment but a mere poke.
due to shaky hand, ive lay down my scalpel for good 4 months now for a ghetto long-blunt 16-18g and a diaper clip, specially for tall no pour agar cups after i saw it cut down my satellite contam rates due to the small surface area of the needles.

I like how they heat up on torch & i can lay a piece of wedge(1/4 size of grain rice) dead center in 2-3 sec without fully lifting the lids of cups..the tissue is protected inside the needle until the plunger(clip) is pushed.

Edited by pacmanbreed (01/14/18 03:27 PM)

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: pacmanbreed] * 1
    #24913525 -

Ingenious. :rockon:
Everybody has to adapt to their particular circumstances and find ways of doing things that work for them.


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

PHENO HUNTERS

Edited by Josex (01/14/18 03:19 PM)

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Josex] * 1
    #24913536 -

Wow that's super cool dude :rockon:


--------------------
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: pacmanbreed] * 1
    #24913601 -

pacmanbreed said:



thats hot shit dude. i have shaky hands too. i just kinda work with the shakes tho.. :oldman:


--------------------
:hypnotoad:
Making Liquid Culture :  Go insane with LC
Mushboy likes these teks : Making shroom tea
Dank Education

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: mushboy]
    #24913603 -

:awelol:


--------------------
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]
    #24913715 -

Josex said:
I mean, if you are working with a known clean culture there's no need to take biopsies from them to transfer, it's slow and you have to take into account that such a tiny winy piece of tissue needs to undergo a lot of cellular division to start a colony from scratch, so no need to subject the culture to unnecessary stress.


I do understand that with a transfer from a culture, a small biopt/transfer from leading edge gives you a better chance to NOT transfer along a contam. I don't see much difference between a needle biopt or a small transfer, both from the leading edge (let's say rice grain size).

The bacteria present are in smaller numbers with a needle biopt and the mycelium amount is also smaller. IMO it's about the conditions that must favor the myc and at the same time inhibit the bacterial growth. That's the deciding factor. Be it antibiotics, tea or BRF mix.

With a supposed clean culture it is very important IMO to make a small transfer from the leading edge. That's because one minimizes the chance of transferring along a contam that wasn't visible to the naked eye.

But I am clearly talking about different situations here; a culture that is probably clean (looking) and one wants to make a second transfer for security OR a known contaminated culture of which one knows it's probably contaminated all over.... only salvageable with antibiotics, tea or BRF mix.. A bit bigger or smaller transfer doesn't matter much here (IMO, alway's an important addition)

Tell me if I am wrong (I might). This just reflects what I am thinking... Trying to form my own image of how things work...

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Shroomway] * 1
    #24913839 -

Shroomway said:
Josex said:
I mean, if you are working with a known clean culture there's no need to take biopsies from them to transfer, it's slow and you have to take into account that such a tiny winy piece of tissue needs to undergo a lot of cellular division to start a colony from scratch, so no need to subject the culture to unnecessary stress.


I do understand that with a transfer from a culture, a small biopt/transfer from leading edge gives you a better chance to NOT transfer along a contam. I don't see much difference between a needle biopt or a small transfer, both from the leading edge (let's say rice grain size).

The bacteria present are in smaller numbers with a needle biopt and the mycelium amount is also smaller. IMO it's about the conditions that must favor the myc and at the same time inhibit the bacterial growth. That's the deciding factor. Be it antibiotics, tea or BRF mix.



You're correct, it won't matter much if you take a biopsy or a little wedge from the contaminated culture, but only in those instances where there's not a lot of bacteria or when bacteria seems to be growing evenly in the plate. Both wedge or biopsy are going to carry bacteria to the receiving puck in proportion to the size of the sample taken.

But when your dealing with severe bacterial contamination, like having a pitiful myc colony drowning in bacterial gunk like I explained in the OP, targeting those areas where there's myc is much easier with a needle. No need to take a wedge and probably transfer more gunk than myc to the receiving puck.
This tek aims at dealing in a effective way with even the worst case scenario.


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

PHENO HUNTERS

Edited by Josex (01/14/18 05:36 PM)

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Josex]
    #24914920 -

mushboy said:
pacmanbreed said:



thats hot shit dude. i have shaky hands too. i just kinda work with the shakes tho.. :oldman:


Try it mush. Its helps alot with our shaky,manly,hairy arms. :awelol:

Josex i have a concern using this particular pp5 of ours.

Sure They fit snugly and their are water but not quite air tight on the lid specially after few cycles.
And i feel it harbors contam through the nook and cranies specially if not use straight from the pc.


- ive been cling wraping their side avoiding the ge before going to the pc. the cling wraps survives and sticks to them making them almost like a one piece protecting their lid`s underside before use.
- I only remove the wrap inside the sab ready to innoc.

Had omit foil due to side ge
but still observing if cling make a difference.
Need to make controls/test if time permits

Edited by pacmanbreed (01/15/18 04:57 AM)

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: Josex]
    #24914938 -

Josex said:
Shroomway said:
Josex said:
I mean, if you are working with a known clean culture there's no need to take biopsies from them to transfer, it's slow and you have to take into account that such a tiny winy piece of tissue needs to undergo a lot of cellular division to start a colony from scratch, so no need to subject the culture to unnecessary stress.


I do understand that with a transfer from a culture, a small biopt/transfer from leading edge gives you a better chance to NOT transfer along a contam. I don't see much difference between a needle biopt or a small transfer, both from the leading edge (let's say rice grain size).

The bacteria present are in smaller numbers with a needle biopt and the mycelium amount is also smaller. IMO it's about the conditions that must favor the myc and at the same time inhibit the bacterial growth. That's the deciding factor. Be it antibiotics, tea or BRF mix.



You're correct, it won't matter much if you take a biopsy or a little wedge from the contaminated culture, but only in those instances where there's not a lot of bacteria or when bacteria seems to be growing evenly in the plate. Both wedge or biopsy are going to carry bacteria to the receiving puck in proportion to the size of the sample taken.

But when your dealing with severe bacterial contamination, like having a pitiful myc colony drowning in bacterial gunk like I explained in the OP, targeting those areas where there's myc is much easier with a needle. No need to take a wedge and probably transfer more gunk than myc to the receiving puck.
This tek aims at dealing in a effective way with even the worst case scenario.



Yeah, a syringe is definitely a very comfortable way of targeting a specific area. Very much so...

@Offlinepacmanbreed: it might help if, in the future, you don't close any PP5 container. Wrapping them in foil with the lid loosely on top doesn't work? Because then they are able to keep their shape

Edited by Shroomway (01/15/18 05:20 AM)

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: pacmanbreed]
    #24914944 -

I haven't experienced that problem myself, well at least I haven't had a problem with my containers until I ran into an exceptionally severe mold contamination in my house (long story). Only then some unused plates really contaminated after a time (2-3 weeks) while being stored in a plastic tote.
But then again, the glad mini rounds I used for agar also contaminated, and even sooner than my current containers. So in my case, it wasn't really anything to do with the choice of containers but rather something to do with the snap-on lids of pp5 containers in general not being perfect and full proof to begin with.

Although similar, those containers you use are not the same as the ones I use.


The lids of my containers close tight with an audible crack and the same crack can be heard when you open them. Also, it's crucial when choosing deli type containers for no-pour that the plastic be relatively thick and the design as plain and simple as possible, so I'd avoid containers with unnecessary bevels, grooves and raised bottoms.

The problem you're having might have something to do with using lids and containers that haven't had the same number of pc cycles, e.g. a lid that has undergone 6 cycles won't close tight on a cup that has had 3 cycles. So in order to prevent this it's imperative that you always use the same lid for the same cup, that'll make a huge difference and will prolong the life of the container considerably.

I'm writing up a no-pour guide atm and I'm covering all this among many other things. In your case, it shouldn't be a bad idea to protect the lids with cling wrap as you're doing. If you think that's a hassle, you could also just trash those containers and purchase more appropriate ones.
If you live in Europe I can give you a link to the ones I'm using.


--------------------
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]
    #24914985 -

Shroomway said:
@Offlinepacmanbreed: it might help if, in the future, you don't close any PP5 container. Wrapping them in foil with the lid loosely on top doesn't work? Because then they are able to keep their shape


Thanks for the tip brother. Ive added a ge for pressure after i vacuum/implode ruined some of my cup without g.e.

Josex said:
I haven't experienced that problem myself, well at least I haven't had a problem with my containers until I ran into an exceptionally severe mold contamination in my house (long story).


I red that horror story.
Youve almost used a box of packing tape to fix your windows :cool: :crazy2: intant ghetto emergency fix.
love your diligence perseverance & observation.
https://www.shroomery.org/forums/showflat.php/Number/24856706#24856706

Josex said:
snap-on lids of pp5 containers in general not being perfect and full proof to begin with.



Its the reason ive began cling wraping them before pc for later use. 1 week for my few leftover plate with cling on a high spore load area now. Ill try to remove the cling if it will contam.

I can see how thick ur pp are. Looks simply sturdy.
Unfortunately not in europe.

Josex said:
The problem you're having might have something to do with using lids and containers that haven't had the same number of pc cycles, e.g. a lid that has undergone 6 cycles won't close tight on a cup that has had 3 cycles. So in order to prevent this it's imperative that you always use the same lid for the same cup, that'll make a huge difference and will prolong the life of the container considerably.



Spot on brother.

I think its the difference when i began using this new pp5 with attached lid & slightly thicker. Almost 25 cycles now
And still hearing audible crack..

I would defenitely will stalk that write up brother. :thumbup:

Edited by pacmanbreed (01/15/18 06:21 AM)

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: pacmanbreed]
    #24915021 -

@Pacman, would you mind if I use those pics in my write-up of the container that's giving you shit? A pic of the inside of an empty container to show the bottom would be awesome if you don't mind. I'd appreciate it a lot.


--------------------
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]
    #24915324 -

Not at all brother, gladly will do.
Coincidentaly ive just finish washing them all. :tongue:
been loving all the patient writeups like yours & contribution shared here by fellows to make it easy for starter like me.

Just saved the images brother since i may accidentally delete them cleaning my galleries if time comes.
Dont give credit brother.

This are my old ones with swapped untight lid like you mentioned. Has a beveled buttom.




This are my new one flat buttomed, ilike how the lids baffle 3x from outside. Guess i can omit clings.





lovely growth pattern. :heart:
-is it mea or grain?
-how far away xtranfer from ms or clone?

Edited by pacmanbreed (01/15/18 12:12 PM)

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: pacmanbreed]
    #24915366 -

I saved the images too, thanks brother.

Oh, so beveled bottom is the name uh? :lol:
:highfive:


--------------------
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]
    #24915369 -

Punt or "kick up" is the name for the beveled bottom on wine bottles and liquor bottles. No one knows that even English speakers so those are useless

I would just say the raised bottom or convex


--------------------
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My Q&A thread. Ask me questions here rather than in Private Message
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5g a day keeps the 5g away

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Re: The Biopsy Method: No Mercy for Bacteria. [Re: bodhisatta]
    #24915371 -

Thanks! :rockon:


--------------------
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] * 2
    #24915469 -

Fuck ya on the no pour tek:rockon:


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🅃 🄴 🄰 🄼    🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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