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Offlinevanceen
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Re: Flow hood - how long to run before use? [Re: JHOVA]
    #28144737 - 01/17/23 04:38 PM (1 year, 2 months ago)

Quote:

JHOVA said:
Ok that explains it. So essentially youve never done done pf tek yourself 100% and you are now struggling with clean spawn. Well that makes sense.




Thanks, but could you explain? I don't really see the connection between having used pre-sterilized BRF and near 100% contamination of grain jars.

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OnlineBeefSupremeJr
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Re: Flow hood - how long to run before use? [Re: vanceen] * 1
    #28144740 - 01/17/23 04:41 PM (1 year, 2 months ago)

i think JHOVA was talking to thugnar.  sorry thugnar tends to make a mess of threads continually though weve all asked him to stop.  he pulls info out of his ass. hes never used a hood.  you might want to put him on ignore just to make things simpler.

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Offlinevanceen
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Re: Flow hood - how long to run before use? [Re: BeefSupremeJr]
    #28144741 - 01/17/23 04:42 PM (1 year, 2 months ago)

Quote:

Thugnar said:
I would suggest OP that you post these guys a pic of your flowhood and learn to make a better agar solution. Sounds like you made it too thick. Im just starting that stuff so i wont be giving any advice about it even if i could.
Some of them are mean, cranky and a bit anal, but they will help you if you give enough information. Start with the flowhood pic. Sounds sus.
Building your own SAB is a good option also.




Here it is:


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Invisiblebakedbeings
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Re: Flow hood - how long to run before use? [Re: BeefSupremeJr] * 1
    #28144758 - 01/17/23 04:49 PM (1 year, 2 months ago)

well here we are...yet another fucking thread gone to shit. this is, as you say, a bad look for the shroomery. here is the exchange thudnar is referring to. as you can see, he is interpreting it in the way most convenient to his ego:

Quote:

Thugnar said:
Ive been told that there is new findings that make some of the "old" techniques "outdated". For example could someone show me the experiements that contradict RR's methods of spawning to bulk and colonizing in high co2 conditions with GE trough the imperfect lid?
Also i would like to understand how pouring boiling water over coir is better than learning proper pasteurizing techniques right from the start? Using the oven and thermometers seems super essential to me in mycology.
Ive been told to use the search engine for many times but the fact is that ive used it for 12 years and even today i can not find these things. If it is true that this information i have is "outdated" then ofcourse i want to learn the new stuff. To me it just seems like there is very little of it here in mushcult and most of the things used today are the same things that were known to work in the beginning of the 20th century. Help would be appreciated.



Quote:

Crackatoa said:
I can't say that I can show you the experiments but through the years people have done side by side grows of straight to fruiting and having the consolidation period. It's been shown to produce fruits faster with the same, if not better results. Just like coir is contaminant resistant so there's no need for proper pasteurization unless you are using things like manure or coffee that will go bad very quickly. The bucket method took off and now people are using room temp water to hydrate their coir. I still believe that myc prefers a "cooked" coir over a straight from the tap hydrated sub. Mush cult has came a long way since RR's info with people trying new things. When I started nobody grew Pans in mono, it was unheard of and now that's a thing. Hopefully this helps.



then....
Quote:

Thugnar said:
Well i already talked to a TC and the conclusion was basicly that nothing ground breaking in growing cubes has been discovered here in mushcult in 10 years... Growing Pan's better and some agar stuff but most of the "new" stuff is just old yarn in new clothing.








OP, my advice for you is also to do PF from scratch. start there, if you succeed move on. cleaning your home will not improve your sterile technique

Edited by bakedbeings (01/17/23 05:05 PM)

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OfflineNuclearTidalWave
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Re: Flow hood - how long to run before use? [Re: BeefSupremeJr]
    #28144763 - 01/17/23 04:51 PM (1 year, 2 months ago)

Quote:

BeefSupremeJr said:
i think JHOVA was talking to thugnar.  sorry thugnar tends to make a mess of threads continually though weve all asked him to stop.  he pulls info out of his ass. hes never used a hood.  you might want to put him on ignore just to make things simpler.




I think JHOVA was actually talking to OP here. Thungar does make it difficult by introducing his advice that clearly lacks first hand experience.

Basically OP has never done PF Tek from the ground up the way it was meant to. By buying pre-made BRF jars you're skipping a lot of important steps hoping that they were all done correctly.

OP your contam issues are likely from buying crap pre-made jars. Just make them yourself it's really not that hard and by doing it yourself you know it was done right.

Edit: I apologize for assisting in the derailment of another thread. I won't add anything further from the peanut gallery.


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Edited by NuclearTidalWave (01/17/23 05:08 PM)

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InvisibleMycolorado
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Re: Flow hood - how long to run before use? [Re: vanceen]
    #28144777 - 01/17/23 05:01 PM (1 year, 2 months ago)

Quote:

vanceen said:
Quote:

Mycolorado said:
How are you inoculating the agar and grain jars?




The agar is inoculated by spores, LC, and by agar to agar transfer.

The jars are mostly LC (after testing the LC on agar), sometimes agar to grain.




Glad you’re testing those LCs!  It’s possible that your culture work needs some improvement.  Are your spores coming from prints or syringe?  Are your LCs started with agar or spores?  When you do agar to grain, are they pristine cultures?  What’s your lid setup on grain jars.  I’m just inclined to lean in that direction of the process as opposed to jumping to the conclusion that your hood is faulty, though I have no clue if their hoods are good or not.

I know it’s been said but wouldn’t trust purchased plates.  You could leave one untouched and see if anything grows on it.  Once you get your own plates going, you could open some in front of the hood and let them sit a while then seal up to test the efficacy of the hood itself. 

Edited by Mycolorado (01/17/23 07:00 PM)

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Invisibleaintlifegrand
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Re: Flow hood - how long to run before use? [Re: vanceen]
    #28144937 - 01/17/23 06:15 PM (1 year, 2 months ago)

OP do you live at high altitude?  Are you venting your PC?  Rinsing your grain after soak?  Bumping up pc time to 2 hours wouldn't hurt

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Offlinevanceen
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Re: Flow hood - how long to run before use? [Re: NuclearTidalWave]
    #28145149 - 01/17/23 09:27 PM (1 year, 2 months ago)

Quote:

NuclearTidalWave said:
Quote:

BeefSupremeJr said:
i think JHOVA was talking to thugnar.  sorry thugnar tends to make a mess of threads continually though weve all asked him to stop.  he pulls info out of his ass. hes never used a hood.  you might want to put him on ignore just to make things simpler.




...

Basically OP has never done PF Tek from the ground up the way it was meant to. By buying pre-made BRF jars you're skipping a lot of important steps hoping that they were all done correctly.

OP your contam issues are likely from buying crap pre-made jars. Just make them yourself it's really not that hard and by doing it yourself you know it was done right.

...






Thanks. I did use purchased jars at the beginning, but most of what I have done has been with jars I made myself. (The exception being the BRF jars.)

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Offlinevanceen
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Re: Flow hood - how long to run before use? [Re: bakedbeings]
    #28145154 - 01/17/23 09:33 PM (1 year, 2 months ago)

Quote:

bakedbeings said:
...
OP, my advice for you is also to do PF from scratch. start there, if you succeed move on. cleaning your home will not improve your sterile technique




Thanks. I actually got about six dry grams using PF tek (purchased jars), but that was from only two jars out of two batches of eight each.

I'm actually looking for ways to improve my sterile technique. I described it in my first post, and that seems to conform with everything I've read.

The only things I've seen recently that I might add are wiping down things rather than just wetting them with isopropyl, and using bleach to kill trichoderma spores on surfaces. I'll report back on whether that helps.

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Offlinevanceen
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Re: Flow hood - how long to run before use? [Re: Mycolorado]
    #28145164 - 01/17/23 09:43 PM (1 year, 2 months ago)

Quote:

NuclearTidalWave said:
Quote:

BeefSupremeJr said:
i think JHOVA was talking to thugnar.  sorry thugnar tends to make a mess of threads continually though weve all asked him to stop.  he pulls info out of his ass. hes never used a hood.  you might want to put him on ignore just to make things simpler.




...

Basically OP has never done PF Tek from the ground up the way it was meant to. By buying pre-made BRF jars you're skipping a lot of important steps hoping that they were all done correctly.

OP your contam issues are likely from buying crap pre-made jars. Just make them yourself it's really not that hard and by doing it yourself you know it was done right.

...






Thanks. I did use purchased jars at the beginning, but most of what I have done has been with jars I made myself. (The exception being the BRF jars.)
Quote:

Mycolorado said:
Quote:

vanceen said:
Quote:

Mycolorado said:
How are you inoculating the agar and grain jars?




The agar is inoculated by spores, LC, and by agar to agar transfer.

The jars are mostly LC (after testing the LC on agar), sometimes agar to grain.




Glad you’re testing those LCs!  It’s possible that your culture work needs some improvement.  Are your spores coming from prints or syringe?  Are your LCs started with agar or spores?  When you do agar to grain, are they pristine cultures?  What’s your lid setup on grain jars.  I’m just inclined to lean in that direction of the process as opposed to jumping to the conclusion that your hood is faulty, though I have no clue if their hoods are good or not.

I know it’s been said but wouldn’t trust purchased plates.  You could leave one untouched and see if anything grows on it.  Once you get your own plates going, you could open some in front of the hood and let them sit a while then seal up to test the efficacy of the hood itself. 




I've made LC both from spores and from mycellium grown on agar (sluicing a slice of colonized agar with sterilized water in a syringe and then pulling it ito the syringe to inject into the LC).

My spores have all come from syringes.

The agar to grain transfers come from what look like pristine cultures. However, I haven't done that too many times. I have mostly used LC for grain.

My lids are mostly two part metal lids with filters for gas exchange and self healing injection ports. I keep micromembrane tape over both of those.

I understand the concern about purchased plates, but I've kept uninoculated plates around for as long as a couple of months with no signs of anything growing in them. That seems like a good sign to me. As I said, I've had better luck with plates than anything else, about 80% have grown a significant amount of white mycellium with no detectable contamination. But I'll give making my own another try.

Thanks for your input.

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Offlinevanceen
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Re: Flow hood - how long to run before use? [Re: aintlifegrand]
    #28145168 - 01/17/23 09:46 PM (1 year, 2 months ago)

Quote:

aintlifegrand said:
OP do you live at high altitude?  Are you venting your PC?  Rinsing your grain after soak?  Bumping up pc time to 2 hours wouldn't hurt




No, I'm in Houston, at about 80 feet above sea level. I let the PC vent for at least five minutes before putting the vent on.

If the contamination is in the grain before inoculation, shouldn't I expect to see contaminants growing in uninoculated jars? I've had some around for quite a while and I've never seen any contamination until a couple of weeks after inoculation.

Thanks for your suggestions.

EDIT: Yes, I do rinse the grain after soaking.

Edited by vanceen (01/17/23 09:50 PM)

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Invisibleaintlifegrand
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Re: Flow hood - how long to run before use? [Re: vanceen]
    #28145183 - 01/17/23 10:06 PM (1 year, 2 months ago)

Quote:

vanceen said:
Quote:

aintlifegrand said:
OP do you live at high altitude?  Are you venting your PC?  Rinsing your grain after soak?  Bumping up pc time to 2 hours wouldn't hurt




No, I'm in Houston, at about 80 feet above sea level. I let the PC vent for at least five minutes before putting the vent on.

If the contamination is in the grain before inoculation, shouldn't I expect to see contaminants growing in uninoculated jars? I've had some around for quite a while and I've never seen any contamination until a couple of weeks after inoculation.

Thanks for your suggestions.

EDIT: Yes, I do rinse the grain after soaking.




90 minutes should kill everything but possible bacteria so you probably wouldn't notice until you got some mycelium growth.  It's worth a try my grain is not good at 90 minutes.

It sounds like something is up with your flow hood.  You could try making an SAB it would be a $15 experiment to rule out the hood.  Just keep in mind contaminants get places with gravity (or flow with your hood) and you can develop technique easy enough

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Re: Flow hood - how long to run before use? [Re: vanceen] * 2
    #28145184 - 01/17/23 10:09 PM (1 year, 2 months ago)

Take a step back and post your agar plates and do agar to grain first. If its healthy spawn it.

Then once you have shoeboxes rolling try grain to grain.

Mess with liquids a lot later once your agar is perfect. LC can go bad fast if the agar work isnt perfect.

Theres a progression to more advanced techniques you gotta go through or at least thats whats recommended.

I like having cakes going at all times as backup. Cakes should be 95% success so theres room for improvement there. You really cant fuck up spores to pf tek jars. Its practically fool proof even in open air which i do all the time.

Do you have pics of your current agar? What about your jars?

Try using a sab as mentioned before. I think that might help since at least you know that works whereas your filter or fan might need tweaking or be mismatched.


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Offlinevanceen
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Re: Flow hood - how long to run before use? [Re: JHOVA]
    #28145547 - 01/18/23 07:36 AM (1 year, 2 months ago)

Thanks all.

I started off using an SAB. That's what I did the PF Tek stuff with.

I'll post some agar plates and failed jars later.

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Offlinevanceen
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Re: Flow hood - how long to run before use? [Re: aintlifegrand]
    #28145877 - 01/18/23 12:41 PM (1 year, 2 months ago)

Quote:

aintlifegrand said:
Quote:

vanceen said:
Quote:

aintlifegrand said:
OP do you live at high altitude?  Are you venting your PC?  Rinsing your grain after soak?  Bumping up pc time to 2 hours wouldn't hurt




No, I'm in Houston, at about 80 feet above sea level. I let the PC vent for at least five minutes before putting the vent on.

If the contamination is in the grain before inoculation, shouldn't I expect to see contaminants growing in uninoculated jars? I've had some around for quite a while and I've never seen any contamination until a couple of weeks after inoculation.

Thanks for your suggestions.

EDIT: Yes, I do rinse the grain after soaking.




90 minutes should kill everything but possible bacteria so you probably wouldn't notice until you got some mycelium growth.  It's worth a try my grain is not good at 90 minutes.

It sounds like something is up with your flow hood.  You could try making an SAB it would be a $15 experiment to rule out the hood.  Just keep in mind contaminants get places with gravity (or flow with your hood) and you can develop technique easy enough




I'll try two hours. Thanks.

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OfflineBobgas
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Re: Flow hood - how long to run before use? [Re: vanceen]
    #28146010 - 01/18/23 02:18 PM (1 year, 2 months ago)

Couple of thoughts:

Quote:

vanceen said:
I tried to post this before but it never appeared. Sincere apologies if this is a duplicate.

To make a long story short, I've been trying to cultivate for a little over six months, with a dismal success rate.

I've had a little success (2 jars out of 16) with PF tek (both from spores and LC), but a 0% success rate with grain in jars or bags. Typically, the mycellium starts to grow and seems to do fine for a week or two, then growth stalls and everything goes green. (Presumably trichoderma.)

Stick with PF Tek

I've done OK with agar dishes, whether from spores or LC. Maybe about 80% success rate, with bacteria, trichoderma, or cobweb mold knocking out about 20%.

Your success rate ought to be much higher

I've really put a lot into trying to create and work in a sterile environment. I've got a 10'x10' room, a room filter running 24/7, I fog the room with disinfectant spray before working, and my workspace and everything that goes into it is sprayed and/or wiped with 70% isopropyl. Syringe needles and scalpels are heated to red flame before use. I'm working in front of a small (12") flow hood, I'd bet a hefty sum this is the issue. 12" is tiny and there is a very small sweet spot of clean flow to work in. the one that Midwest sells with the acrylic extensions coming forward from the sides.

Reading around, I've found that it's recommended to start up the vent hood an hour or two before working. This I have not done, I always start it up right before use.

Here's the question: Would not running the flow hood an hour or two before working be expected to lead to very high contamination rates?

I even saw a 2mm worm living inside one of my agar dishes wtf a couple of days after I had inoculated it! The only thing I can think of is that the flow hood is blowing crap, including worm eggs, during the first hour or two. Does this make sense to you all?




Make an SAB or splurge for a 2X2 FFU (or build a FH if that's your thing - following a proven design.)  The other stuff you are doing is unnecessary unless your environment requires it.

6 months of this and not having success must be exceedingly frustrating!

Head to the PF tek thread and follow it exactly.  Do the work yourself.  Get spores from a trusted vendor. Fruit your successful cakes in a water tub.

Vent your pc for 10 -15 mins


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Offlinevanceen
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Re: Flow hood - how long to run before use? [Re: Bobgas]
    #28146060 - 01/18/23 02:47 PM (1 year, 2 months ago)

Quote:

Bobgas said:
Couple of thoughts:

Quote:

vanceen said:
I tried to post this before but it never appeared. Sincere apologies if this is a duplicate.

To make a long story short, I've been trying to cultivate for a little over six months, with a dismal success rate.

I've had a little success (2 jars out of 16) with PF tek (both from spores and LC), but a 0% success rate with grain in jars or bags. Typically, the mycellium starts to grow and seems to do fine for a week or two, then growth stalls and everything goes green. (Presumably trichoderma.)

Stick with PF Tek

I've done OK with agar dishes, whether from spores or LC. Maybe about 80% success rate, with bacteria, trichoderma, or cobweb mold knocking out about 20%.

Your success rate ought to be much higher

I've really put a lot into trying to create and work in a sterile environment. I've got a 10'x10' room, a room filter running 24/7, I fog the room with disinfectant spray before working, and my workspace and everything that goes into it is sprayed and/or wiped with 70% isopropyl. Syringe needles and scalpels are heated to red flame before use. I'm working in front of a small (12") flow hood, I'd bet a hefty sum this is the issue. 12" is tiny and there is a very small sweet spot of clean flow to work in. the one that Midwest sells with the acrylic extensions coming forward from the sides.

Reading around, I've found that it's recommended to start up the vent hood an hour or two before working. This I have not done, I always start it up right before use.

Here's the question: Would not running the flow hood an hour or two before working be expected to lead to very high contamination rates?

I even saw a 2mm worm living inside one of my agar dishes wtf a couple of days after I had inoculated it! The only thing I can think of is that the flow hood is blowing crap, including worm eggs, during the first hour or two. Does this make sense to you all?




Make an SAB or splurge for a 2X2 FFU (or build a FH if that's your thing - following a proven design.)  The other stuff you are doing is unnecessary unless your environment requires it.

6 months of this and not having success must be exceedingly frustrating!

Head to the PF tek thread and follow it exactly.  Do the work yourself.  Get spores from a trusted vendor. Fruit your successful cakes in a water tub.

Vent your pc for 10 -15 mins




I appreciate your response. Yes, it is a frustrating. And yes, seeing that worm was a WTF moment for sure.

As I've mentioned above, I've got an SAB. I haven't used it except for the PF tek, and I had dismal results with that, even though I followed the recommendations closely. Basically the same story; stalled mycellium growth, then green takes over.

Two questions, if I may:

1. When you say "the other stuff" I'm doing is unnecessary, specifically which actions do you mean?

2. This is the first I've heard of fruiting PF in a water tube. I have always dunked the cakes overnight, rolled in vermiculite, then fruited them in a tub containing field capacity perlite. Do you mean to actually place the cakes in water within a tub?

Thanks again.

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Invisiblebakedbeings
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Re: Flow hood - how long to run before use? [Re: vanceen]
    #28146078 - 01/18/23 02:58 PM (1 year, 2 months ago)

cakes on a rack suspended over an inch of water, or in their jars sitting in the water, or on foil boats sitting on saturated coir

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Offlinevanceen
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Re: Flow hood - how long to run before use? [Re: vanceen]
    #28146138 - 01/18/23 03:29 PM (1 year, 2 months ago)

In case anyone's interested, I did a quick check of the airflow in front of my vent hood using an alcohol flame.

The flame pointed straight away from the surface of the filter, both close in and up to a foot or so away. The flame was steady.

I did see some fluttering of the flame near the corners of the filter, and of course it fluttered a lot as soon as I moved it out of the area of the filter.

This isn't rigorous, but it seems like good news. (Although it doesn't solve my problem.)

Rather than jumping into building a bigger hood, I'm going to try some experiments with agar, e.g. leaving one dish open for a defined time away from the hood and in front of it. I'll probably also try one as soon as I turn the motor on, then maybe a half hour and an hour later.

Any comments or suggestions would be appreciated. You've all helped me out a good bit.

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Re: Flow hood - how long to run before use? [Re: vanceen] * 2
    #28146156 - 01/18/23 03:35 PM (1 year, 2 months ago)

Yep, as I mentioned earlier, I’d test the hood with some plates open in front of it so you can rule it out as the problem.  Honestly, I would say it’s your technique/purchased plates and grain that are the root of your issues.

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