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OfflineTheProdigy
People like youjust fuel myfire...

Registered: 12/10/04
Posts: 164
Last seen: 14 years, 8 months
Cambodian Contamination Problem =(
    #4097348 - 04/25/05 11:42 PM (16 years, 5 months ago)

I'm on my 3rd batch of cambodians. I made this batch with some Creeper (Same substrate, same Pressure Cooker, etc.) and out of 3 batches of cambodians, this time I have a single surviving jar. The rest have fallen victim to the blue-green mold (The creeper are doing completely fine). I'd give pics, but everyone who could probably help me on this probably knows what blue green mold looks like (Or at least I'm assuming they do).

I PC for 45 mintues to 60 minutes at 15 PSI, I use ~ 1/4 BRF to Verm combo, and the rest is history. Can anyone think of anything?

-The Prodigy


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

If you haven't gone to http://www.boohbah.com while you are tripping... it's a must.


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Offlinescrname
Stranger

Registered: 03/09/05
Posts: 196
Last seen: 1 year, 3 months
Re: Cambodian Contamination Problem =( [Re: TheProdigy]
    #4097364 - 04/25/05 11:45 PM (16 years, 5 months ago)

Glovebox?


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Offlinejroe
Stranger
Registered: 03/31/04
Posts: 39
Last seen: 15 years, 7 months
Re: Cambodian Contamination Problem =( [Re: TheProdigy]
    #4097371 - 04/25/05 11:47 PM (16 years, 5 months ago)

hmm... try several of these ideas that i have been thinking about....

**use lysol to steralize the air, specially before fanning or anything during hte casing...

**use the myco enviro bag for the inocculation. almost 100% contamination free... then you can transfer to a casing

** be anally sterile while making your casing. microwave the hell out of it, lysol the air, soap your hands ect....

other ideas?
make the condition bad for anything outside the jar, casing box , chamber ect. what i mean is... keep the rest of the room at a low humidity level to stop external bacteria growth. this is if your casing/jars can have proper humidity inside their area though


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OfflineTheProdigy
People like youjust fuel myfire...

Registered: 12/10/04
Posts: 164
Last seen: 14 years, 8 months
Re: Cambodian Contamination Problem =( [Re: scrname]
    #4097377 - 04/25/05 11:48 PM (16 years, 5 months ago)

I'm not anal enough about contams to use a glovebox yet, although I would like to have one, I thought about it already, would probably solve my problem though.


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

If you haven't gone to http://www.boohbah.com while you are tripping... it's a must.


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OfflineTheProdigy
People like youjust fuel myfire...

Registered: 12/10/04
Posts: 164
Last seen: 14 years, 8 months
Re: Cambodian Contamination Problem =( [Re: TheProdigy]
    #4097381 - 04/25/05 11:50 PM (16 years, 5 months ago)

Quote:

**use lysol to steralize the air, specially before fanning or anything during hte casing...




This is a *really* good idea, but won't I be looking at some possible mutants? (I have nothing against handicapped mushrooms, but let's face it, they are ugly!)


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

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Offlinepsilocyben
bad guy

Registered: 12/03/04
Posts: 1,026
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Re: Cambodian Contamination Problem =( [Re: scrname]
    #4097384 - 04/25/05 11:51 PM (16 years, 5 months ago)

maybe you can better identify your contaminant  here
if it's penicillium, i can assure you that you don't want that laying around!:dead:
when you get a chance, look at some of the symptoms of the mycotoxins in some common contaminants  here
"I PC for 45 mintues to 60 minutes at 15 PSI,"
it could not hurt to try 90 minutes.
are they spores from a previous grow?
if so, ever had any problem with contamination?


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




Edited by psilocyben (04/25/05 11:55 PM)


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OfflineAbermelin
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Registered: 03/22/05
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Re: Cambodian Contamination Problem =( [Re: TheProdigy]
    #4097393 - 04/25/05 11:53 PM (16 years, 5 months ago)

theirs alot of variables involved that you didnt mention.

First thing i can tell you is that your PC'ing too long. Biologists are taught to only PC something 15psi for 15 minutes to sterilize, although most shroomers are taught for 45 which is excessive, 30 mins is good. you could be drying out your cakes which hinders mycellium growth, im not sure what effect that has on contam growth.

either way, make sure that any holes in your lids arent exposed, do this by putting a layer of tinfoil over the lid and tape/polyfill over the holes. make sure the seal on your lid is good if your using jar lids.

as long as your following the innoc PF procedure, i dont see why you should be having any problems. you are flaming the needle after every insertion into the jar right?

if your doing everything right, i recommend getting a hepa filter. also, throw some desiccant in a container in your innoc room a few days before hand, it helps lower mold concentration in the air.

more likely than not, your innoc conditions are bad. you are one big contam.


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Offlinepsilocyben
bad guy

Registered: 12/03/04
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Re: Cambodian Contamination Problem =( [Re: Abermelin]
    #4097404 - 04/25/05 11:58 PM (16 years, 5 months ago)

Quote:

Abermelin said:
Biologists are taught to only PC something 15psi for 15 minutes to sterilize, although most shroomers are taught for 45 which is excessive, 30 mins is good.





could you show me info supporting this abermelin?
reason i ask is that i have heard otherwise for some time and i'm curious what you're source is.


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OfflineTheProdigy
People like youjust fuel myfire...

Registered: 12/10/04
Posts: 164
Last seen: 14 years, 8 months
Re: Cambodian Contamination Problem =( [Re: Abermelin]
    #4097424 - 04/26/05 12:03 AM (16 years, 5 months ago)

Actually this does look a *lot* like penicillium... especially in this pic they presented in an example. It completely fits the description, too. http://www.shroomery.org/images/23418/green2.jpg

Give me a little bit and I will post a picture.

Let me answer the details that Abermellin posed, first, though.

Quote:

First thing i can tell you is that your PC'ing too long. Biologists are taught to only PC something 15psi for 15 minutes to sterilize, although most shroomers are taught for 45 which is excessive, 30 mins is good. you could be drying out your cakes which hinders mycellium growth, im not sure what effect that has on contam growth.




I can say for 1, that the jars are definitely not dehydrated. Not to sound too blunt about that, but they are not getting dehydrated.

Quote:

either way, make sure that any holes in your lids arent exposed, do this by putting a layer of tinfoil over the lid and tape/polyfill over the holes. make sure the seal on your lid is good if your using jar lids.




I follow the PF tek almost by the book. I inoculate through a double layer of tin foil, and then place a layer of sterile foil over the top of that (All pressure cooked with the jars)

Quote:

you are flaming the needle after every insertion into the jar right?




No, but I do flame it till red after every other insertion to a different jar. The PF tek calls for it every 3 jars, so I feel I'm being fairly good about it.

-The Prodigy


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

If you haven't gone to http://www.boohbah.com while you are tripping... it's a must.


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Offlinepsilocyben
bad guy

Registered: 12/03/04
Posts: 1,026
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Re: Cambodian Contamination Problem =( [Re: TheProdigy]
    #4097459 - 04/26/05 12:12 AM (16 years, 5 months ago)

do you wipe it down with alcohol between each inoculation?
if not, this is most likely your problem.
did that myself one time.

i'm serious about that penicillium, though.
GET IT OUT!
it's sporulation is airborne and ubiquitious(fucking everywhere)
you could risk permanent healt risks including
respiratory and neurological damage.
NASTY STUFF! (no more smeling)


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




Edited by psilocyben (04/26/05 12:15 AM)


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OfflineTheProdigy
People like youjust fuel myfire...

Registered: 12/10/04
Posts: 164
Last seen: 14 years, 8 months
Re: Cambodian Contamination Problem =( [Re: TheProdigy]
    #4097461 - 04/26/05 12:13 AM (16 years, 5 months ago)

Here's a pic of my problem. It's not but in the early stages of development, but it looks a lot texture and color wise to the picture that I presented from the examples. It also has an ammonia/vinnegar type smell to it.



Enjoy!


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

If you haven't gone to http://www.boohbah.com while you are tripping... it's a must.


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OfflineTheProdigy
People like youjust fuel myfire...

Registered: 12/10/04
Posts: 164
Last seen: 14 years, 8 months
Re: Cambodian Contamination Problem =( [Re: TheProdigy]
    #4097468 - 04/26/05 12:15 AM (16 years, 5 months ago)

Quote:

do you wipe it down with alcohol between each inoculation?




Wipe what down, the needle? Nope. I use fire to sterilize the tip.


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

If you haven't gone to http://www.boohbah.com while you are tripping... it's a must.


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Offlinepsilocyben
bad guy

Registered: 12/03/04
Posts: 1,026
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Re: Cambodian Contamination Problem =( [Re: TheProdigy]
    #4097502 - 04/26/05 12:22 AM (16 years, 5 months ago)

get a clean alcohol soaked paper towel to wipe the entire needle down.
use flame when you begin to kill anything inside the needle, then cool it with the towel.
this almost surely your problem, i'm afraid.

with those jars-
fill the sink up with 50/50 bleach and water.
submerge jar
open, and let it sit under water for the better part of an hour to kill your contaminants.
dispose and then soak your jars in a 10% bleach water sol. for 12 hours
dishwash
start over and remember your experience.

that's not the one good jar is it?


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




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OfflineTheProdigy
People like youjust fuel myfire...

Registered: 12/10/04
Posts: 164
Last seen: 14 years, 8 months
Re: Cambodian Contamination Problem =( [Re: psilocyben]
    #4097555 - 04/26/05 12:35 AM (16 years, 5 months ago)

no, that's not the good jar. The jars that are surviving are thriving, rhizomorphic mycelium. I can't wait till they are all grown up *tear*. Too bad only one camb jar made it though. I'm excited for the creeper, however.

Will Peroxide work as opposed to bleach or alcohol, you think? I'm going to get some lysol next chance I get as well.


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

If you haven't gone to http://www.boohbah.com while you are tripping... it's a must.


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Offlinepsilocyben
bad guy

Registered: 12/03/04
Posts: 1,026
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Re: Cambodian Contamination Problem =( [Re: TheProdigy]
    #4097574 - 04/26/05 12:40 AM (16 years, 5 months ago)

no dice on the peroxide.
i'm sure you could use alcohol, too, but i stopped using that for the disenfection of equipment some time ago as it is not practical.

great as a surface cleaner in a spray bottle though.

agar reccomends the use of white vinegar and hydrogen peroxide, separately.
spray one, wait a moment, then spray with the other.
he said it will kill ANYTHING.


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




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Offline_JJ_
PsychedelicRevolutionary
Registered: 06/12/01
Posts: 369
Loc: NSW, Australia
Last seen: 6 years, 3 months
Re: Cambodian Contamination Problem =( [Re: psilocyben]
    #4097583 - 04/26/05 12:44 AM (16 years, 5 months ago)

Did you make up your own syringes? If you did and you didn't use a glove box I would say that's where your problem lies.


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OfflineTheProdigy
People like youjust fuel myfire...

Registered: 12/10/04
Posts: 164
Last seen: 14 years, 8 months
Re: Cambodian Contamination Problem =( [Re: psilocyben]
    #4097587 - 04/26/05 12:44 AM (16 years, 5 months ago)

Thanks for the input man. =)


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

If you haven't gone to http://www.boohbah.com while you are tripping... it's a must.


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Offlinestarseed1066
officially hosed

Registered: 04/13/05
Posts: 325
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Re: Cambodian Contamination Problem =( [Re: TheProdigy]
    #4097607 - 04/26/05 12:50 AM (16 years, 5 months ago)

Quote:

TheProdigy said:
I'm not anal enough about contams to use a glovebox yet, although I would like to have one, I thought about it already, would probably solve my problem though.




i have not used lysol once. no pressure cooker, no H2O2, and no glovebox.

but if you do, just wash your hands, pay close attention to everything you touch, and whether or not any sterile surfaces touch any non-sterile surfaces.

0% contamination i promise.


--------------------
Under his instruction, I taped drumsticks to his head and turned him into a sort of mummy, or perhaps a caterpiller in a cocoon. He remains this way for about three hours, making bizarre noises, pretending to be a new species that must learn to walk and talk and eat, etc. And he communicates with other life forms by way of the antennae on his head. Eventually we jam a tube from a waterbong into it's mouth and figure he's learned all he needs to know.


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Offlinewhatever123
Whatever I did, I'm sorry
Male User Gallery

Registered: 04/07/05
Posts: 2,613
Loc: San Diego, CA
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Re: Cambodian Contamination Problem =( [Re: _JJ_]
    #4097628 - 04/26/05 12:57 AM (16 years, 5 months ago)

Quote:

_JJ_ said:
Did you make up your own syringes? If you did and you didn't use a glove box I would say that's where your problem lies.





Wow....That is a damned good point. Did you make your own?


--------------------
Koala Koolio said:
there should be a 3 month waiting period between registration and posting. :wink:


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OfflineTheProdigy
People like youjust fuel myfire...

Registered: 12/10/04
Posts: 164
Last seen: 14 years, 8 months
Re: Cambodian Contamination Problem =( [Re: whatever123]
    #4097807 - 04/26/05 02:01 AM (16 years, 5 months ago)

Actually, I did not make these syringes, I wish I could say I did and that is where the problem is, but nope, not it.


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

If you haven't gone to http://www.boohbah.com while you are tripping... it's a must.


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OfflineAbermelin
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Registered: 03/22/05
Posts: 394
Loc: In The Mycelial Network
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Re: Cambodian Contamination Problem =( [Re: psilocyben]
    #4097865 - 04/26/05 02:26 AM (16 years, 5 months ago)

Quote:

psilocyben said:
Quote:

Abermelin said:
Biologists are taught to only PC something 15psi for 15 minutes to sterilize, although most shroomers are taught for 45 which is excessive, 30 mins is good.





could you show me info supporting this abermelin?
reason i ask is that i have heard otherwise for some time and i'm curious what you're source is.




i have this file called lab_techniques.pdf that was posted in an older thread. Its a biologists guide to sterilization and other fungi related processes. you can search it, i dont know the exact thread. it gives a chart of the correlation between PSI and cooking time.


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InvisibleSoopaX
Criminal DrugAnalyst

Registered: 11/13/04
Posts: 1,690
Re: Cambodian Contamination Problem =( [Re: Abermelin]
    #4097927 - 04/26/05 05:04 AM (16 years, 5 months ago)

Dude, stop spreading your misinformation.


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


Jackie Treehorn treats objects like women, man


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Offline_JJ_
PsychedelicRevolutionary
Registered: 06/12/01
Posts: 369
Loc: NSW, Australia
Last seen: 6 years, 3 months
Re: Cambodian Contamination Problem =( [Re: SoopaX]
    #4097954 - 04/26/05 06:01 AM (16 years, 5 months ago)

"In a laboratory, items are sterilised in autoclaves at 121?C for 15 minutes or at 126?C for 10 minutes. The times mentioned here are average sterilisation times. The time that objects and materials are autoclaved depends on their size as large objects take longer to sterilise. Also, the autoclave should be loosely packed so that the steam can circulate around the objects and so heat them to the correct temperature."
- From here.

The key is it depends on the size of the object being sterilized. A couple of quart jars of rye grain will take longer than some pf jars. It will take longer to heat the center of the jar to the same temperature as the outside if the jar is larger.
15-20 minutes @ 15psi should be fine for a batch of PF jars in my opinion, but if you're doing quarts 30-40 mins is a better idea. No point saving yourself ten to fifteen minutes when your whole grow can be ruined.


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OfflinePhotoguy
Drunk andJobless

Registered: 02/24/05
Posts: 553
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Re: Cambodian Contamination Problem =( [Re: _JJ_]
    #4098004 - 04/26/05 07:46 AM (16 years, 5 months ago)

I'm not trying to argue here, but I PC for 90 minutes, and if my PC is all full, i generally will do a boil of the other jars for a minimum of 2.5 hours. I haven't had the need to build a glove box yet, although I'm sure i should.

I highly doubt that PC'ing for a long time will alter your water content that much as to hinder your growth. Hell, i was drunk and PC'd one night, let it cool down, the next evening i got drunk and PC'd the same jars again. Well, those jars are thriving so far, and frankly this has been the best grow, because I have had zero contams show up. Who knows??

I would just say this, do the jars for 90 minutes. Whats the big freakin deal you impatient people, its going to take you several weeks to get through a grow anyways, whats a lil more time?


--------------------
You gotta check yo' self before you wreck yo' self

B, Z, F, Burma, Cambodian, Golden Teacher, Puerto Rico, South American, Costa Rica, African

My name is Ender Wiggen and I will own you soon. Bean is my second in command, he is PC'ing jars right now.


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Offlinepsilocyben
bad guy

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Re: Cambodian Contamination Problem =( [Re: _JJ_]
    #4098190 - 04/26/05 10:33 AM (16 years, 5 months ago)

Quote:

_JJ_ said:
The key is it depends on the size of the object being sterilized.




agreed;
the link provided was probably speaking in regards to agar plates or something.


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




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OfflineAbermelin
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Re: Cambodian Contamination Problem =( [Re: psilocyben]
    #4098358 - 04/26/05 11:53 AM (16 years, 5 months ago)

Quote:

_JJ_ said:
The key is it depends on the size of the object being sterilized.





makes sense. you prob would have to increase times if your were PCing a quart jar of sub, but i would go by the listed psi/times when doing half pints, as they are not large or thick enough to need excessive PCing, and i dont think the starter of this thread is doing quart brf jars. once you get past a certain length of time, your just cooking your jars.

but as far as over PCing goes, i used to have problems with the top of the cake drying out with brf/verm cakes, as that type of substrate holds moisture poorly, but this could have been attributed to air exchange. but whatever, if you encounter this problem, go with grains.


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InvisibleSemilanceata
No god, no boss

Registered: 05/26/03
Posts: 841
Loc: República Federal Íbera
Re: Cambodian Contamination Problem =( [Re: TheProdigy]
    #4098387 - 04/26/05 12:10 PM (16 years, 5 months ago)

If you prepared your cambodian jars same way as the creepers, and inoc. the same way, I would say your print/syringe is contamed.


--------------------
Sr_Setahongo


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Offlinepsilocyben
bad guy

Registered: 12/03/04
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Re: Cambodian Contamination Problem =( [Re: Semilanceata]
    #4098398 - 04/26/05 12:13 PM (16 years, 5 months ago)

he only sterilized his needle once before, and once during with flame. no alcohol at all.

it's likely that this was his problem.

if not from a previous grow,
was the source of the spores a user or a vendor?


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




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InvisibleSemilanceata
No god, no boss

Registered: 05/26/03
Posts: 841
Loc: República Federal Íbera
Re: Cambodian Contamination Problem =( [Re: psilocyben]
    #4098424 - 04/26/05 12:26 PM (16 years, 5 months ago)

I thought he said the needle gets flamed till red hot. Anyway, if a scalpel, inoc.loop or needle gets flame sterilized, why wipe some alcohol afterwards? Isn?t fire the best sterilizer?


--------------------
Sr_Setahongo


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Offlinepsilocyben
bad guy

Registered: 12/03/04
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Re: Cambodian Contamination Problem =( [Re: Semilanceata]
    #4098439 - 04/26/05 12:34 PM (16 years, 5 months ago)

first, to cool it.
if you were to flame-sterilize inbetween each inoculation point,
think about the amount of time you'd waste waiting for the needle to cool each time. hardly practical.
if no flowhood or glove-box to innoc in,
then you don't want that needle sitting in open air for a few minutes while it cools before it goes in the jar.

i had contamination problems ONE time. and that was when the needle was wiped down between jars, and not between each inoculation site.
i always flame sterilize when i begin to kill anything inside the needle.


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




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Offline_JJ_
PsychedelicRevolutionary
Registered: 06/12/01
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Last seen: 6 years, 3 months
Re: Cambodian Contamination Problem =( [Re: psilocyben]
    #4100077 - 04/26/05 08:20 PM (16 years, 5 months ago)

I don't see why you would need to wait for it to cool.. just insert it a couple of seconds later, it'll cool down the second you start ejecting spore solution (or starts melting the polyfill :P) Really though, I wait about 2-3 seconds and the it does not damage the polyfill at all.


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Offlineoysterguy63
60's child

Registered: 04/08/05
Posts: 463
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Re: Cambodian Contamination Problem =( [Re: _JJ_]
    #4100561 - 04/26/05 10:51 PM (16 years, 5 months ago)

I just did very first grow with Cambo's, but utilized cakes as a rookie. Had six cakes on perlite in an aquarium with a submersible heater and a stone to provide some oxygen....temp at 74 F. Got my substrate from pfjars and all six just whooped ass on the first flush.



--------------------
Oysters...yuummmmm


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Offlinepsilocyben
bad guy

Registered: 12/03/04
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Re: Cambodian Contamination Problem =( [Re: oysterguy63]
    #4100990 - 04/27/05 01:34 AM (16 years, 5 months ago)

i imagine that your spore solution might cool down the needle,
but isn't it also going to heat up your soluution as it comes out.

obviously, it has worked for you, right?
just trying to be objective and make sure everyone, (including both you and i) learns the most efficient way of doing things.


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




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Offlinestarseed1066
officially hosed

Registered: 04/13/05
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Re: Cambodian Contamination Problem =( [Re: psilocyben]
    #4101208 - 04/27/05 02:21 AM (16 years, 5 months ago)

i havent had a problem with heat sterilizing the needle and injecting right away... the first drop or two might be pretty hot but the subsequent 1ml is definitely not too hot to cause problems.


--------------------
Under his instruction, I taped drumsticks to his head and turned him into a sort of mummy, or perhaps a caterpiller in a cocoon. He remains this way for about three hours, making bizarre noises, pretending to be a new species that must learn to walk and talk and eat, etc. And he communicates with other life forms by way of the antennae on his head. Eventually we jam a tube from a waterbong into it's mouth and figure he's learned all he needs to know.


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* Post deleted by users_request Uv1 2,548 11 03/03/18 02:14 PM
by bodhisatta
* Corn On The Cobweb, ContamiNation KingCobWeb 1,860 7 11/24/02 05:50 AM
by KingCobWeb
* Re: don't throw out those contaminated jars... JoHnNySc 7,849 9 12/14/99 12:21 AM
by Psilowarrior
* problem bigwussy 736 3 08/26/02 12:36 AM
by XAZIA
* Invitro Contamination with chronic tek MDP1182 2,187 3 07/29/02 06:28 AM
by bigslick

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