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Invisibleturtlelamp
n00b
Registered: 11/27/03
Posts: 93
Loc: The Midwest
Now we're up to five consecutive generations of no-shows...
    #2184033 - 12/16/03 09:26 PM (20 years, 3 months ago)

What with the search function disabled, I can't reference my last post on this, but essentially, I have not been able to get a single jar to even germinate the last five times I've prepared jars. I came home today five days after innoculating 8 brf jars expecting to see SOMETHING in ONE of my jars. I even tried a new vender, sporeworks, and innoculated the jars with the potent looking syringes not five minutes after Fedex came to my door. Still nothing. And I know it's not impatience, since every time I've had success in the past I've seen growth after three days. What could I possibly be doing wrong? SO wrong that not ONE jar will even germinate? Even if I microwaved the syringes and injected them into my ass 8 times, surely I'd get more than I'm getting now. I'm at a total loss. One thing I have noticed is that last time, and now this time too, the brf has darkened almost imperceptibly directly under the innoculation points. Pretty much exactly where the solution runs down the side of the glass. Bacterial? Possibly. They don't smell bad. They smell pretty strongly of over-cooked brown rice flour. I just can't figure it out. Help! I'm not ready to give up yet, but I'm getting closer.

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Offlinemobiusrunner
myco maniac
Registered: 10/13/03
Posts: 129
Last seen: 14 years, 30 days
Re: Now we're up to five consecutive generations of no-shows [Re: turtlelamp]
    #2184041 - 12/16/03 09:33 PM (20 years, 3 months ago)

1.) How long have you waited?
2.) What conditions are you keeping the jars in?
3.) Are you sure it's organic BRF? I've heard of some non-organic BRF having fungicide and/or other nasties in it.


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By "I", I of course mean a good friend of mine who happens to live in the fine city of Amsterdam. "I" is just so less awkward.

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Offlineprotocoldroid
synthtek aa
Registered: 07/21/01
Posts: 122
Loc: central ny
Last seen: 19 years, 10 months
Re: Now we're up to five consecutive generations of no-shows [Re: turtlelamp]
    #2184044 - 12/16/03 09:34 PM (20 years, 3 months ago)

this might be completely off track, but...

i was colonizing some KS's on WBS, and... I thought they'd colonize really fast (well compared to the PF cubies i'm used to), but, waited about a week, and nothing there. so i check the temperature, and krikes, they're at 65 deg. I get the heat up to about 78 and bam, day later i've got mycelium.

also, check your ratio of ingredients in your BRF... you might have too much / too little water.

on growing boomers in your ass... well, if works, write a tek, "TP tek". peace!


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-=/ "what is this electrofunk that's drivin' y'all crazy?" \=-
[disclaimer: a science-fiction post for a science-fiction world, this post is not intended to be true.]

Edited by protocoldroid (12/16/03 09:36 PM)

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Invisibleturtlelamp
n00b
Registered: 11/27/03
Posts: 93
Loc: The Midwest
Re: Now we're up to five consecutive generations of no-shows [Re: mobiusrunner]
    #2184056 - 12/16/03 09:41 PM (20 years, 3 months ago)

Quote:

mobiusrunner said:
1.) How long have you waited?



It's been five days (I know, I know...) but I've seen it so many times before. There's no fungal life in those jars.

Quote:


2.) What conditions are you keeping the jars in?




82F dual-rubbermaid with 40% rh.

Quote:


3.) Are you sure it's organic BRF? I've heard of some non-organic BRF having fungicide and/or other nasties in it.



It's Bob's Red Mill. I got it at my local, ritzy, Jewish, whole foods store.

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InvisibleGoldenShroom
shhhh ....don'ttell anyone!

Registered: 03/13/03
Posts: 91
Loc: wherever shrooms grow wil...
Re: Now we're up to five consecutive generations of no-shows [Re: turtlelamp]
    #2184061 - 12/16/03 09:45 PM (20 years, 3 months ago)

Pics of the jars would be of great help.... Otherwise, the typical answer would be that the water content is off.


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-The killer in me is the killer in you... so I send this smile over to you

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Invisibleturtlelamp
n00b
Registered: 11/27/03
Posts: 93
Loc: The Midwest
Re: Now we're up to five consecutive generations of no-shows [Re: GoldenShroom]
    #2184096 - 12/16/03 10:07 PM (20 years, 3 months ago)

Here they are: 1/4 cup of water, 1/4 cup of brf, 2/3 cup of verm



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InvisibleHanky
wiffle bat.
Male User Gallery
Registered: 08/30/03
Posts: 56,993
Loc: Great Southern Land.
Re: Now we're up to five consecutive generations of no-shows [Re: turtlelamp]
    #2184110 - 12/16/03 10:13 PM (20 years, 3 months ago)

try a different brand of BRF,i cant think of anything else that would cause consistant faliures.
are you PCing your jars or steaming?


--------------------
Coaster is an idiot...
[quote]Coaster said:
but i thnk everything thats pure is white?
[/quote]



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Invisibleturtlelamp
n00b
Registered: 11/27/03
Posts: 93
Loc: The Midwest
Re: Now we're up to five consecutive generations of no-shows [Re: Hanky]
    #2184362 - 12/17/03 12:33 AM (20 years, 3 months ago)

Does anyone think they look too wet or too dry? I was under the impression that less water is better, because they'll colonized faster. But I can still see tiny droplets on the glass and the brf is moist, so I think it's just about right.


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Fingerprints on file.

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OfflinePsilocybeingzz
Male User Gallery

Registered: 12/15/02
Posts: 14,463
Loc: International waters
Last seen: 11 years, 4 months
Re: Now we're up to five consecutive generations of no-shows... [Re: turtlelamp]
    #2184377 - 12/17/03 12:44 AM (20 years, 3 months ago)

5 days is nothign I have had jars take longer then the so called "2 week limit of no return". :smile:

If there is nothing in 3 weeks open up ONE and see what went on inside!, did it stall???? or not grow at all

you DO TAKE OFF your tape right??????(after pcing???)


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Invisibleeric_the_redS
Trans-male User Gallery

Registered: 02/28/03
Posts: 14,475
Loc: happy land
Re: Now we're up to five consecutive generations of no-shows [Re: turtlelamp]
    #2184378 - 12/17/03 12:45 AM (20 years, 3 months ago)

looks like you have too much verm. in there.

try using only 1/2 cup instead of 2/3 cup next time.
incubate them at 84 degrees and you should have no problem.


--------------------
Anno cock? is that some kind of Greek liqueur? -Geo's All Knowing Sex Slave

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Invisibletheseeker
addict
 User Gallery
Registered: 12/27/01
Posts: 489
Re: Now we're up to five consecutive generations of no-shows... [Re: turtlelamp]
    #2184464 - 12/17/03 01:32 AM (20 years, 3 months ago)

possibly overcooked? you can get away with lower pressure and 45 min-1 hour for BRF. I also agree with OOKLA about having the rubber side up.

some other ideas, hope they help.
* is verm:brf:water in 2:1:1?
* brf has no preservatives?
* incubating at what temp? is it consistent?
* incubating at ok RH?
* jars allowed to cool before inoc?
* syringe water allowed to cool before xfer spores?
* no alcohol/fresh h2o2 making it into the syringe?
* jar bands on too tight/too loose?
* brf compacted?

open a jar up after cooling. see if it's too dry, too wet, or overcooked. if it is, buy some cheap cracked corn, boil kernels for 30 mins, fill jars 2/3 and pc @ 15psi for 70 mins. good luck with the brf though!

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InvisibleMagash
Da Bud Guru
 User Gallery

Registered: 07/25/02
Posts: 5,876
Loc: Near Hilo
Trusted Cultivator
Re: Now we're up to five consecutive generations of no-shows... [Re: turtlelamp]
    #2185212 - 12/17/03 12:08 PM (20 years, 3 months ago)

This is gonna sound stupid as fuck but somewhere here I read that you had success before and all of a sudden it stopped. What are you washing the jars with? A friend was using dish soap that is antibacterial (which I recamend) but was leaving a residue or whatever and when he used regular soap his problems stopped. I haven't had the same problem but it is something to look into.

The name of the stuff in the soap was called triclosan . That's the chemical that kills spores.


--------------------
All creatures tremble when faced with violence. All creatures fear death, all love life. If we can only see ourselves in others, then how could we possibly hurt another creature?


:growingweed: Join us at the Growery! :growingweed:

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Invisibleturtlelamp
n00b
Registered: 11/27/03
Posts: 93
Loc: The Midwest
Re: Now we're up to five consecutive generations of no-shows... [Re: Magash]
    #2185283 - 12/17/03 12:51 PM (20 years, 3 months ago)

That's actually a really good point. Maybe a contributing factor? The thing is, I did have success my first time. The second time, when the failures began, however, I was using entirely new jars (since the first were still colonizing). It wasn't until after that that I started actually washing jars. At first I was washing them with anti-bacterial soap before preparing the jars... but now I've taken to soaking them in a bleach solution for a few days before just sticking them in the dishwasher. I don't believe the dishwasher soap has any anti-bacterial agents... but that's something I'll be double checking as soon as I get home. You must be on to something, though. Because SOMETHING is destroying these spores. They are arriving to me viable, and somewhere along the way they are getting fucked up. Last time the jars cooled two days. It's not that. The syringes looked good... I'm a little concerned because ups left the spores on my porch the morning they delivered them (it's cold in my neck of the woods) but they didn't freeze, and from what I understand, spores are tough. So I doubt it's that. I'm confused. Next time I prepare jars I'm not even going to pc them. Yeah, they'll likely get contaminated, but I don't even care about that at this point. I just want to see some mycelium.


--------------------
Fingerprints on file.

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Offlineforevrgrounded98
Stranger

Registered: 10/25/03
Posts: 283
Last seen: 17 years, 1 month
Re: Now we're up to five consecutive generations of no-shows... [Re: turtlelamp]
    #2185339 - 12/17/03 01:24 PM (20 years, 3 months ago)

you could boil them in water or put em in the oven or flame sterilize em after you wash them, i know how dissapointing it is to see no growth after like 3 weeks.
-matt


--------------------
"If I don't see ya before the end of this one, I'll meet ya in the next one and don't be late" - Jimi Hendrix

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Offlinedjd586
Underpants Gnome

Registered: 02/03/03
Posts: 1,655
Last seen: 14 years, 1 month
Re: Now we're up to five consecutive generations of no-shows... [Re: turtlelamp]
    #2185421 - 12/17/03 01:54 PM (20 years, 3 months ago)

In the past, I never took time to wash new 1/4 pint jars. I just figured that if there were any contams on or in the jars that they would be killed durning the sterilization process. My sucess rate was always at or near 100%.

Then I started to read that everyone was presterilizing their jars. I thought that it couldn't hurt so I also started to presterilize. I wiped the jars down with rubbing alchohol, let them sit in a bleach wash for a few hours(1:50, bleach to water), washed them out with some dish soap and water, then I rinsed the jars out well with deionized water. After that, I followed my normal routine for a PF tek style BRF cake. Incubated my jars for 2 weeks and only got growth in one jar. 1 out of 60! I thought for sure I got some bum syringes. I contacted my vendor, explained the situation, and they were nice enough to resupply me.

When my new syringes arrived, I decided not to presterilize my jars... followed my same old routine. Innoculated 60 jars, incubated, seen growth in 4 days in most jars. In 24 days, 57 of 60 cakes were completely colonized. The other three took an extra week. But none-the-less, I was back to 100% success!

Now I can't be sure that it was the fact that I pre cleaned my jars that lead to my failure. It could have just as easily been that I recieved some bad syringes. Many people presterilize their jars and have great success. ANYHOW, the moral of my post is... try not presterilizing... maybe that's your problem.


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

Phase 1... collect underpants... phase 2...??? ... Phase 3 - PROFIT!

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Invisibleturtlelamp
n00b
Registered: 11/27/03
Posts: 93
Loc: The Midwest
Re: Now we're up to five consecutive generations of no-shows... [Re: djd586]
    #2185469 - 12/17/03 02:09 PM (20 years, 3 months ago)

That's a really good suggestion. I'm going to combine the last two posts, and try ONLY boiling the jars to pre-sterilize. There must be some bleach fumes in there or something that's destroying my spores... nothing else could account for the consistent failures I've had. Thanks for your help, everyone.

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InvisibleLateForTheFuture
Old Hand
I'm a teapot User Gallery

Registered: 02/24/03
Posts: 845
Re: Now we're up to five consecutive generations of no-shows [Re: turtlelamp]
    #2185545 - 12/17/03 02:50 PM (20 years, 3 months ago)

I pre clean my jars every time and still have 100% success, so I do not see this as a factor. I usually run them through the dishwasher and give them another good handwash and set them upside down to dry.  Use this for your mixture, instead of what you were:

1/2 cup verm
1/4 cup BRF
I never have actually gotten a set amount of water I use, but its usually around 1/4-1/2 cup. I just get it to the right consistency (a drop of water falls out if I squeeze the verm)

Also, try mixing your verm and water FIRST before you add the BRF, it seems to mix alot better.

Anyway, don't give up my friend. You might want to try ordering your own print and making a THICK ass syringe just to up your odds. Good luck! :laugh:

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Offlineforevrgrounded98
Stranger

Registered: 10/25/03
Posts: 283
Last seen: 17 years, 1 month
Re: Now we're up to five consecutive generations of no-shows [Re: LateForTheFuture]
    #2185651 - 12/17/03 03:47 PM (20 years, 3 months ago)

Just out of curiosity would adding verm to ur jars make them colonize faster? I never add verm to mine and they generally tend to


--------------------
"If I don't see ya before the end of this one, I'll meet ya in the next one and don't be late" - Jimi Hendrix

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Offlinedjd586
Underpants Gnome

Registered: 02/03/03
Posts: 1,655
Last seen: 14 years, 1 month
Re: Now we're up to five consecutive generations of no-shows [Re: forevrgrounded98]
    #2185734 - 12/17/03 04:24 PM (20 years, 3 months ago)

Yes... Vermicuilte allows for more gas exchange. More O2 in the jars the faster colonization goes.


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

Phase 1... collect underpants... phase 2...??? ... Phase 3 - PROFIT!

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InvisibleHanky
wiffle bat.
Male User Gallery
Registered: 08/30/03
Posts: 56,993
Loc: Great Southern Land.
Re: Now we're up to five consecutive generations of no-shows [Re: LateForTheFuture]
    #2186445 - 12/17/03 08:49 PM (20 years, 3 months ago)

Quote:

LateForTheFuture said:


Also, try mixing your verm and water FIRST before you add the BRF, it seems to mix alot better.






i agree ....this makes a huge difference,the brf will coat the verm if added last,if added first it will clump.


--------------------
Coaster is an idiot...
[quote]Coaster said:
but i thnk everything thats pure is white?
[/quote]



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