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InvisibleLord Toranaga
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Re: First pins in ten years [Re: VaeVictum]
    #14906724 - 08/11/11 04:22 PM (10 years, 2 months ago)

I've been reading a lot about taking prints, and my understanding is that without lab conditions from noc to print, there is no way to capture a 100% sterile specimen.  In thinking so much about sterility, I realize that even bringing the cake from the FC in one room to the bathroom which is another means exposure to unwelcome invaders.

I read that very carefully taken prints probably still have some contaminants present, and one should not over-fill spore solutions in order to avoid transferring a critical amount of them, and ruining the batch.

So I understand that I should be as sterile as possible, and I did do my homework (and still am).  But I still don't know how sterile is enough for prints.

1- Can I bring the cake through open air to my bathroom?
2- Should I always wear latex gloves through the printing process?
3- Can I work without my glovebox if I ousted the crap out of the bathroom and alcohol wiped my tools?
4- If I accidentally drop and touch the top of the cap, should I quit the attempt and dry the fruit instead?

I started using a sterile nail pushed up through the cap to hold onto, but I often feel like my fingers get too close to the cap when its inserted.  In general, this procedure still feels new and awkward, and I know its partly a matter of getting comfortable through practice.  However, any tips would be appreciated.

:cheers:
Quote:

VaeVictum said:
Nice treasure trove of mushies you got going on!



:ahahaha:Little bit o this, little bit o that... love my treasure trove!:mushdance:


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InvisibleVaeVictum
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Re: First pins in ten years [Re: Lord Toranaga]
    #14907274 - 08/11/11 06:18 PM (10 years, 2 months ago)

I've made 3 syringes now. All 3 I made by setting the cap in my 1/2 pint sterilized jar wrapped in foil or you can put the lid on. I then just squirted 11 cc's of boiling water that I sucked up and let cool in there and scraped some print off the bottom and sucked it up then repeated.

I've made 20 BRF jars since and none have contaminated. :shrug: I could just be getting lucky though.

I carry my cake all over the house and haven't had any issues since cakes are so contam resistant once 100%. However though I was told when I was first starting out that the kitchen and the bathroom are the worst places to do anything and it really does make sense after thinking about it.

GL on the prints though, I'm sure you'll figure out a sterile tek to work for you. I've got some cambo/panama prints I could trade ya some time if you want new strains.  :pm:


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Offlinejust me
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Re: First pins in ten years [Re: VaeVictum]
    #14907721 - 08/11/11 07:50 PM (10 years, 2 months ago)

dont worry about printing mang!
at least if its a print youd send me :lol:
the beauties of agar
and ms

printing is so easy tho
cut caps pre harvest(if a tub with tight parameters)
lay a strip of clean foil
put caps on it
cover them with cups(too much humidity imo) or a sweater box type container like ppl use for fruiting.
you could get real fancy and make it filtered and breathable
let the print sit 12-24 hours
lay another clean piece of foil
fold edges
make cuts
fold remaining edges
100.99% sterile! everytime!

and just how Vaevictum said, prints are easy as pie
i do slightly different, but we all do, so :shrug:
i like to fill the jars
to desired amount. about 15cc if filling one 12ml syringe
or 100cc if im filling about 10 ms syringes :lol:
i used to use foil only, but ive found lids and rings not screwed tight are much easier to work with in the glove box...
especially with a small inoculation hole near the rim
and then PC. usually while PC'ing agar, or sterilizing tools because they all take the same amount of time in the PC
let cool
move to glovebox with prints and scalpel
remove lid
scrape print
put lid back on
stir and suck up with you clean syringe
(instead of stirring, i like to let the spores sit in the water until ive made all my MS jars, then stir/swirl them. this allows the spores to hydrate a little, and makes them mix with the water easier, and fewer stick to the sides when you swirl it)
flame needle tip
cap it
label it
date it

yumm pie


--------------------

--------------------------------------------------

-pEaCeLoVeGoDbLeSs-

"The Downfall of Mankind; is Believing He Has Limitations."


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InvisibleVaeVictum
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Re: First pins in ten years [Re: just me]
    #14907728 - 08/11/11 07:52 PM (10 years, 2 months ago)

Forgot to mention I cycle the water in and out of my print jar 2-3 times to disperse all of the spores.

His method sounds a lot more safe than mine.


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Offlinejust me
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Re: First pins in ten years [Re: just me]
    #14907736 - 08/11/11 07:54 PM (10 years, 2 months ago)

i stopped wearing gloves ages ago
i dont remember why
and a few times ive tried going back...
i dont print in my glovebox, although i probably should
i just make sure my area is extremely clean
take the prints, and set something sterile over the top to keep out air born contams
if i were you, id worry less about sterile procedure in printing, and focus on sterile procedure with agar and dishes :grin:


--------------------

--------------------------------------------------

-pEaCeLoVeGoDbLeSs-

"The Downfall of Mankind; is Believing He Has Limitations."


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Offlinescatmanrav
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Re: First pins in ten years [Re: just me]
    #14907825 - 08/11/11 08:22 PM (10 years, 2 months ago)

When growing in a greenhouse right before the veil tears, I move the substrate into a bag or a tub, sealed off, maybe some polyfil. The veils tear within hours, wipe the cap clean with an alcohol napkin, slice it off and print as mentioned, usually I have the foil in a tubberware container with the lid not on tight. I dont use a glove box though just quickly pop the cap into the container.

You can also take a cap thats getting ready to for the veil to tear, and use a sterile utensil to tear it yourself. All the contamination that would end up on the print is usually on the gills, or sometimes the top of the cap. As long as you keep those two areas clean, and what you print on clean during the print, the print is usually pretty clean. Most prints arent 100% clean though.


--------------------
"life is like a drop of rain getting closer and closer to falling into a lake, and then when you hit the lake there is no more rain drop, only the lake."

Growing with bags, start to finish (including my new grain and substrate prep)
Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
How I do grain (old still good tips)
Turn your closet into a fruiting chamber
Casing layer colonization and overlay


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Offlinenikesb1949
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Re: First pins in ten years [Re: VaeVictum]
    #14907850 - 08/11/11 08:28 PM (10 years, 2 months ago)

Should have B+ spores in about 2 weeks. Would love to trade for cambo


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OfflineImmortal Corrupter
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Re: First pins in ten years [Re: nikesb1949]
    #14908031 - 08/11/11 09:06 PM (10 years, 2 months ago)

I did it Shirley Knot's way, just wiping the tinfoil down with alcohol and putting a pushpin in the cap to pick up, works fine for me.

I also use an inoculation loop I made from a bread tie. Light bread tie on fire, make loop, done, and worked great streaking the spores onto my agar :smile:


--------------------
where's my lap dance?
.:nonono:
/OO\
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What poor gods we do make


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InvisibleLord Toranaga
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Re: First pins in ten years [Re: just me]
    #14908370 - 08/11/11 10:23 PM (10 years, 2 months ago)

@Immortal Corrupter, scatmanrav, just me, and VaeVictum:
Thanks guys, always an abundance of good, freely flowing info round these parts.:thumbup:
I'll probably do a little of what each of you said, but in general, I just wanted to hear:
Quote:

just me said:
dont worry about printing mang!
at least if its a print youd send me :lol:
the beauties of agar
and ms
if i were you, id worry less about sterile procedure in printing, and focus on sterile procedure with agar and dishes :grin:




I'll generally be quite clean and sterile, but now I also understand that who gets the print and what teks they use affects how sterile their usable prints need to be.  This inspires me that much further into agar, because I figure I'll end up with more viable prints with agar techniques.

:hi5:

Too bad I missed the few hour window that the homebrew store is open.  I'll have to wait for tomorrow, again..:picard:


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InvisibleVaeVictum
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Re: First pins in ten years [Re: Lord Toranaga]
    #14908385 - 08/11/11 10:27 PM (10 years, 2 months ago)

You sir...are....now aboard...the...

:failboat:

Just kidding, get there tomorrow in time though!

You'll never get to work on agar at this rate.


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InvisibleLord Toranaga
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Re: First pins in ten years [Re: VaeVictum]
    #14908436 - 08/11/11 10:35 PM (10 years, 2 months ago)

Quote:

VaeVictum said:
You sir...are....now aboard...the...

:failboat:

Just kidding, get there tomorrow in time though!

You'll never get to work on agar at this rate.




...took me a minute. I was like, what's he saying..? then :lolsy:


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InvisibleLord Toranaga
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Re: First pins in ten years [Re: Lord Toranaga]
    #14913027 - 08/12/11 09:59 PM (10 years, 2 months ago)

Ya'll are not gonna believe this shite, but I got the pics to prove it...

Those of you who have been watching this post for a minute will recall that I had a false contam alarm and trashed a tray of upcoming Creeper beauties.  Well...  I first put the cut up cake in its own plastic grocery bag in the (mostly empty)trash, but then I thought I might as well try to plant it outside somewhere.  That was three days ago (page 4), and I ended up just stashing the Creepy grocery bag inside a different clean trash bag on my kitchen floor.

So tonight, I was stirring some baked beans on the stove, and I looked down at the trash bag on the floor, and I had this no-fucking-way moment where I suddenly had to dig into the bag and smell if there was any chance that the bulk substrate was still occupied solely by p.cubes.

To my herculean amazement the bag smelled totally fresh and cubey, with no sour or sweet odors!!!!!  I just about crapped my pants, and proceeded to immediately initiate Operation Frankenstein.  Here's the result:

Am I totally crazy?  Will this inevitably doom the rest of the martha to contams?  Feel free to give helpful advice, or just stay tuned for the saga :wink:

For people like me who can't get enough:
The second R44 cake is getting mature... ...as its bigger brother bulks for the second, similarly top-heavy second flush
The first Peru cake also is plumping its second flush, always beating the R44 in weight through numbers


Edited by Lord Toranaga (08/12/11 10:22 PM)


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Invisiblelostinbq
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Re: First pins in ten years [Re: Lord Toranaga]
    #14913136 - 08/12/11 10:30 PM (10 years, 2 months ago)

i really dont kno if i should be happy about that pinset or in fear it may come alive and eat me while i sleep.. cakes look good tho.


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InvisibleLord Toranaga
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Re: First pins in ten years [Re: lostinbq]
    #14913289 - 08/12/11 11:06 PM (10 years, 2 months ago)

Quote:

lostinbq said:
or in fear it may come alive and eat me while i sleep



:feelsbadman:
Hopefully Its Monstrosity will forgive my bad judgment, and be happy it gets another chance to live..?  ...I hope ( :aweneck: )


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OfflinePrimalSoup
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Re: First pins in ten years [Re: Lord Toranaga]
    #14913425 - 08/12/11 11:37 PM (10 years, 2 months ago)

Quote:

immediately initiate Operation Frankenstein.  Here's the result:




You're one lucky bastard, that's all.:laugh2:  Looka dem pins! :mushroomgrow:

:peace:PS


--------------------


if you stand too close to the machine it'll start to eat you
Primal's simple tested teks and projects: :awesomenod: Wheat Prep 2.0  Acidic Tea Tek  Potency Project! 


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Offlinejust me
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Re: First pins in ten years [Re: PrimalSoup]
    #14913586 - 08/13/11 12:16 AM (10 years, 2 months ago)




Am I totally crazy?



:aweneck:

just dont let nasty shit sporulate all over


--------------------

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-pEaCeLoVeGoDbLeSs-

"The Downfall of Mankind; is Believing He Has Limitations."


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InvisibleLord Toranaga
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Re: First pins in ten years [Re: just me]
    #14913639 - 08/13/11 12:27 AM (10 years, 2 months ago)

Quote:

just me said:
just dont let nasty shit sporulate all over




Yeah, I'll keep a close eye, and figure I'd get some amount of visible warning before total mushroom armageddon. Its also at the bottom of the martha, on a longer chain.

Yessir.


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Offlinejust me
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Re: First pins in ten years [Re: Lord Toranaga]
    #14913647 - 08/13/11 12:30 AM (10 years, 2 months ago)

tighter leash? longer chain?


--------------------

--------------------------------------------------

-pEaCeLoVeGoDbLeSs-

"The Downfall of Mankind; is Believing He Has Limitations."


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Invisiblerjreynoldsinc
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Re: First pins in ten years [Re: Lord Toranaga]
    #14913656 - 08/13/11 12:32 AM (10 years, 2 months ago)

glad to see your back in the game


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InvisibleLord Toranaga
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Re: First pins in ten years [Re: just me]
    #14913667 - 08/13/11 12:35 AM (10 years, 2 months ago)

Quote:

just me said:
tighter leash? longer chain?



heehee, the poor Creeps deserve a bigger pen, a longer leash...

but I meant that the other shelves in the martha are closer together, and the bottom one hangs real low.  I figured it would be good to keep the possible contams on the bottom and further away from the other trays.


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