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slugworth
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Registered: 09/30/13
Posts: 37
Last seen: 10 years, 1 month
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failing beginner.need help. quitting not option 1
#19066463 - 10/31/13 09:55 PM (10 years, 2 months ago) |
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Hey guys looking for some guidance here. You can review my thread history for complete details but in short I'm having an unanticipated amount of trouble finding success in this hobby. I am definitely discouraged but I'm determined to figure this out. I am a details type person and have no troubles with cultivation skills working in other hobbies. I got three syringes from a vendor back in September. Using brf and the pftek I have nocced up around 16 jars And one honey lc. Sterilized two attempts with regular pot for 90 minutes with two layers of foil over lids (approx. Half of jars have also had tape over the innoc holes during sterilization) and another two attempts with a pc for 60 minutes. In my first batch I had maybe 4/8 jars show Some thIck white rhIzo Growth however all jars were over run with a whiteish grey mold or bacteria. Since then, three attempts later I have essentially wasted two more syringes. The second syringe produced absolutely nothing. No mold bacteria or mycelIum - zero. So far the third syringe is proving no better after 15 days. 3/5 jar show white fuzzy, what I believe is cobweb mold, and two show again absolutely nothing. In each jar the spot starts exactly at the inocculation sites. I have used a glove box with flame sterilization between jars. I have assessed some variables. With the first batch I used 10ml for 8 250 ml jars. With this attempt all jars showed growth however only a few showed rhizomorphIc and all showed white grey contam (odorless). With second attempt I used around 5ml for 5 jars And had zero growth after 25+ days. With third attempt I used just over 1ml per jar and have said cobweb looking stuff in 3/5. I have same brown rice flour pre ground for all attempts, as well as same verm. And tap water - our water is quite chlorinated however I have read on here that Shouldnt be an issue.
I have 5 more syringes coming from a different vendor (first batch did come from a sponsored vendor)
Any words of wisdom, encouragement or help at all for my next endeavor are sought. I can answer any procedural questions for analysis of methods for troublshooting.
I apologize for any weird caps (cap letters that is, I Wish I Could apologize for other weird caps) I'm posting from my phone
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PsiLisaBin
Stranger<<<Strangest


Registered: 07/11/13
Posts: 128
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Re: failing beginner.need help. quitting not option [Re: slugworth]
#19066517 - 10/31/13 10:05 PM (10 years, 2 months ago) |
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my immediate thought here is give up on the LC - just don't even bother with it. a LOT of times the culprit for contamination is a sketchy LC. it's hard to tell just based on sight alone if your LC is clean or not, and if i was a betting man, i'd bet your LC is likely not clean.
just use your spore syringe to inoculate your jars. you're flame sterilizing your needle red-hot between jars, right? get it red hot, then squeeze a drop or two of solution out of it to cool it down, then get to work.
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MisterDeadeye


Registered: 09/02/11
Posts: 171
Last seen: 8 years, 8 months
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Re: failing beginner.need help. quitting not option [Re: slugworth]
#19066529 - 10/31/13 10:06 PM (10 years, 2 months ago) |
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There are different types of mycelium growth. I think it's unlikely that you grew only mold/bacteria in all of your jars.
Do you have any pictures of the jars? Weak mycelium growth can look grey/white. And cobweb is incredibly wispy and would likely take over the entire jar in a couple days. It's very distinct visually.
Edit: Your second batch of jars might have been too hot, killing the spores on contact.
Edited by MisterDeadeye (10/31/13 10:10 PM)
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barong
Nada


Registered: 07/24/11
Posts: 666
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Re: failing beginner.need help. quitting not option [Re: slugworth]
#19066549 - 10/31/13 10:09 PM (10 years, 2 months ago) |
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1. Do you let the PC vent before putting the weight on?
2. Do you shower beforehand, dry of with a newly laundered towel, then put on laundered clothes?
3. Do you wipe your hands down with alcohol?
4. DO you avoid breathing over your work area?
5. Do you spray the air in your work area with an alc/water o bleach/water solution to pull dust particles from the air?
6. DO you work first thing in the morning to avoid dust being kicked up through the day as people move through the house?
There may be no single answer to your issues, just get more fastidious about a clean work area. Without a flowhood it pays to be a bit over-the-top with cleaning and stuff.
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slugworth
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Registered: 09/30/13
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Last seen: 10 years, 1 month
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Re: failing beginner.need help. quitting not option [Re: barong]
#19069552 - 11/01/13 11:55 AM (10 years, 2 months ago) |
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The honey lc was an attempt to preserve the syringes that I had to make them last longer, I haven't nocced any jars with it. As per cooling, with each attempt I give at least overnight or work day to cool - with flame ster. I let it cool around 10-20 secs then squirt some solution through to further cool. Then inject. No alcohol wiping after flame as RR instructed this was counterproductive. Injection setting I used open oven set at 300f the first time. The rest of them have been with a glove box, cleaned with 70% alc and sprayed with lysol generously. I have not showered before each attempt but I have used gloves for 2x and a dust mask in one of those.
One thing I am not certain about is venting the pc ? It has an automated lock mechanism that slips into place once enough steam is blowing off, once that occurs is when I start timing.
I do have pics of the first batch in which there was actually something growing, I can repost those later this evening.
Thanks for input guys. I guess ill continue working steril techniques.
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Stupendous-Yappi
Anomaly XB-311394


Registered: 09/23/13
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Re: failing beginner.need help. quitting not option [Re: slugworth] 1
#19069707 - 11/01/13 12:29 PM (10 years, 2 months ago) |
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Quote:
slugworth said: One thing I am not certain about is venting the pc ? It has an automated lock mechanism that slips into place once enough steam is blowing off, once that occurs is when I start timing.
Doesn't your PC have a pressure gauge and/or regulator? On mine the the locking mechanism engages about ten minutes before the pressure reaches 15 psi. So, you may not be PC'ing for long enough.
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barong
Nada


Registered: 07/24/11
Posts: 666
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Re: failing beginner.need help. quitting not option [Re: slugworth]
#19070278 - 11/01/13 02:50 PM (10 years, 2 months ago) |
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Quote:
slugworth said: I have not showered before each attempt but I have used gloves for 2x and a dust mask in one of those.
You should be aware that the atmosphere we live in is teeming with air borne bacteria, and as you go through your day your body and clothes pick up all of this nasty stuff. I don't have any way of telling you how significant a clean body is but why risk it? Just set everything up the night before, wake up,shower, and do what you have to.
Bathroom towels would be a huge source of bacteria also, as they hang around damp all day. Clean towels, clean clothes.
And a dust mask isn't doing much, the bacteria you're breathing has smaler particals than they can filter. Not that I think it's necessary to wear one, but a surgical mask would probably be better.
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PirateSwazey



Registered: 12/12/12
Posts: 2,993
Loc: Here, Now
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Re: failing beginner.need help. quitting not option [Re: barong]
#19070658 - 11/01/13 04:08 PM (10 years, 2 months ago) |
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post up some pics of your jars so we can look and see what you're calling mold
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slugworth
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Registered: 09/30/13
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Re: failing beginner.need help. quitting not option [Re: PirateSwazey]
#19072092 - 11/01/13 08:44 PM (10 years, 2 months ago) |
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So here's the most recent batch - PC'ed at least 60-90 min. They look quite a bit more "webby" vs the first batches appearance. 
The very first batch which was done using about 1.0cc per jar - they actually showed significant growth after 4 days when I checked em the first time. They still took at least 3 weeks to take over the whole jar. You can see distinct differences in color and texture leading me to believe they were contamed. They do look slightly darker in the pics vs in person and there were no greens, oranges or blues. When I chucked em I did analyze - the texture of the substrate was pretty firm almost like a dried out sponge, quite dense as well, like semi difficult to break apart. There was pretty much no smell, def. nothing foul.
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PirateSwazey



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Posts: 2,993
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Re: failing beginner.need help. quitting not option [Re: slugworth]
#19072172 - 11/01/13 08:59 PM (10 years, 2 months ago) |
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well the new batch looks great...that's exactly what you want. i cant really tell what was up with the other ones. looks like metabolites on some of them but the last 2 look pretty sketchy to me.
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SgtPepperNo9
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Registered: 10/30/13
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Re: failing beginner.need help. quitting not option [Re: slugworth]
#19072179 - 11/01/13 09:01 PM (10 years, 2 months ago) |
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I have no real experience in person as I am on my 1st grow but have looked at a ton of pics and read a ton of stuff. Someone with more experience can answer this better than me and prob correct my post but...since no one has responded yet.
The first set with the 1/2 pint short jars don't look terrible IMO. A little lighter then the jars I have done so far but the outward myc growth looks similar.
The taller 1/2 pints look more concerning to me. Uneven looking growth with some parts looking undisturbed and just covered with grey/light white.
In the 4th pic from the left where you are lifting the cap slightly it looks like the white goes past the lid. Did it cover the dry verm layer too?
Like I said I am sure someone with more experience will respond and you can disregard my opinion.
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JMcDoogle
A Serious Scholar


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Re: failing beginner.need help. quitting not option [Re: slugworth]
#19072253 - 11/01/13 09:14 PM (10 years, 2 months ago) |
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Quote:
slugworth said: So here's the most recent batch - PC'ed at least 60-90 min. They look quite a bit more "webby" vs the first batches appearance. 
The very first batch which was done using about 1.0cc per jar - they actually showed significant growth after 4 days when I checked em the first time. They still took at least 3 weeks to take over the whole jar. You can see distinct differences in color and texture leading me to believe they were contamed. They do look slightly darker in the pics vs in person and there were no greens, oranges or blues. When I chucked em I did analyze - the texture of the substrate was pretty firm almost like a dried out sponge, quite dense as well, like semi difficult to break apart. There was pretty much no smell, def. nothing foul. 
2nd from last picture, looks a bit odd to my noob eyes, but the mycellium growth looks a lot whiter than the growth around it, almost a grey.
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The ego is nothing other than the focus of conscious attention.
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joiningheads
Enthusiast


Registered: 05/05/13
Posts: 41
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Re: failing beginner.need help. quitting not option [Re: slugworth]
#19072254 - 11/01/13 09:15 PM (10 years, 2 months ago) |
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Monokaryotic mycellium looks very similar to cobweb. Thats the mycellium that grows from the individual spore before it finds a compatible mating type to form more rhizo-like growth. Those jars on the top freshly inoculated look fine (for now, but if they aren't properly sterilized bacteria will develop, typically not mold because that only requires ~150F to kill) Based on your jars stalling out constantly and those pics toward the bottom, I'm thinking bacteria is a big problem here. Don't start counting time until the PC reaches 12 or 15 psi which is a good 10-20 minutes after the PC starts pressurizing. Go for a full 90 minutes and for God's sake don't use the oven for a glovebox. The oven tek is bunk; your success rate in the open air will be the same or better than the oven (because the air is moving due to the heat). You want the stillest possible air and don't worry about letting the needle cool just go straight from your alcohol flame to your glovebox with the needle red hot and inject. Work quickly. Wear a surgical mask or have your glovebox setup so you're not breathing bacteria into the work area.
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Stupendous-Yappi
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Registered: 09/23/13
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Re: failing beginner.need help. quitting not option [Re: joiningheads]
#19072354 - 11/01/13 09:30 PM (10 years, 2 months ago) |
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slugworth
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Registered: 09/30/13
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Re: failing beginner.need help. quitting not option [Re: Stupendous-Yappi]
#19082495 - 11/03/13 08:28 PM (10 years, 2 months ago) |
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Well thanks for all the input guys, its definitely encouraging. Since I had already deemed the new batch contaminated I had actually mishandled two of the jars (carried into garage and allowed to tilt at least partially sideways - hopefully that doesn't mess em, but at least theres a few that were undisturbed as well). As for the monokaryotIc growth is that something I should be concerned about ? Is there a definitive rhyme/ reason that I am seeing this ?
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PirateSwazey



Registered: 12/12/12
Posts: 2,993
Loc: Here, Now
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Re: failing beginner.need help. quitting not option [Re: slugworth]
#19085597 - 11/04/13 02:10 PM (10 years, 2 months ago) |
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Quote:
slugworth said: As for the monokaryotIc growth is that something I should be concerned about ? Is there a definitive rhyme/ reason that I am seeing this ?
sometimes the growth is rhizomorphic and sometimes it isn't. i believe it has to do with how much sex the mushrooms are having. not much you can do about it unless you pick up the agar work.
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cronicr



Registered: 08/07/11
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Loc: Van Isle
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Re: failing beginner.need help. quitting not option [Re: PirateSwazey]
#19086416 - 11/04/13 05:26 PM (10 years, 2 months ago) |
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it's a nutrition thing(the diffence between rhizzo and tomentose) and even iso's will act diffrent on diffrent subs, drop colonized graiin on agar and it takes off once it hits the not as nutritious sub
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  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
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Sockadin



Registered: 01/03/10
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Re: failing beginner.need help. quitting not option [Re: cronicr]
#19086555 - 11/04/13 05:56 PM (10 years, 2 months ago) |
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I worry your over thinking this whole thing. Worst case scenario, you might have a few bacteria problems. I mean this in the most productive way possible. The more you mess with those jars, the more problems you will have.
The one thing we haven't talked about is Jar size and Vermiculite barriers. If you are handling them before they have consolidated you might be moving around your "filter"; this is set up on top to block contaminates.
The stalling issue is more than likely because of the wide mouth quart jars that you are using in the bottom set of pictures. I have found that the PF tek works best with either 1/2 pint wide mouth jars, or the 8oz jelly jars.
Put them in a place with light/dark rotations and start your Agar work. They will consolidate and start to do what you want when you leave them be.
Check out my signature for an example of an 8oz Jelly Jar grow. It has a nice timeline as well.
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Enigma1
Positive



Registered: 08/15/13
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Re: failing beginner.need help. quitting not option [Re: Sockadin]
#19086997 - 11/04/13 07:30 PM (10 years, 2 months ago) |
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Are you cutting off all the air flow and ousting the air . Showering a must too. You have a hair net?
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slugworth
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Registered: 09/30/13
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Re: failing beginner.need help. quitting not option [Re: Enigma1]
#19087965 - 11/04/13 11:21 PM (10 years, 2 months ago) |
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First batch was my first attempt ever . Those are the pics up top - lower few in wide mouth jars were attempt 3.5. Was directed by an acquaintance who claimed a degree of experience. I had read of gloveboxes on here but changed my gameplan after he told me that. After further investigation I now believe glovebox in still air room after sterilizing is better, it makes sense . I also did not have a pc although I did place weight on top of my large pot and let steam for 90+. Another mistake I made which is largely debated on here was alcohol wiping after flaming because I wasn't sure how carbon would affect things. Those issues (minus I had two tries with pot before purchasing pc) were corrected in second attempt.
Next hotly debated variable I changed for second attempt (in addition to a new syringe)was ditching the Tit (which was Kept in dark still air location). In the second attempt I waited month and absolutely nothing happened in a single jar. I corrected All of my mistakes, used gloves, had mask on. I even tacked lysol wipes over openings of glove box in case any contaminants tried to drift passed my arms. Legit nothing happened for four weeks. Using same syringe I made an lc which I have yet to test as now I'm being told lc is apparently garbage, recovered my jars (lid, and two foil layers held on buy rubber bands to ensure no entry of excess moisture , pc'ed, re innocced with syringe number two again. Placed in different rooms around house. Again 3+ weeks and not a single jar showed a thing. I consider that attempt 2.5 because I didn't replace the brf mix.
Now I'm onto my third syringe - those are the pics in the bottom. I did 5 jars. Put two in a Tit, this time at a lower temp (78... First time my "experienced acquaintance told me 84...that was before I was educated on the developing myc giving off heat making it warmer and more favorable to bacterial infection which may be what happened in attempt 2 as joiningheads suggested.) The other three I scattered about the house. That is about our current status. When I saw this growth and spent a few more hours reviewing the most widely discussed cobweb threads and pics I thought for sure it was contaminated (despite reading some compelling info by guys that seem to know what they are talking about, stating true cobweb is rare during pftek brf colonization).
As per stalling out, none have technically stalled unless this hobby has a specific definition of the word stalling. First batch was contaminated (used a whole syringe for 8 half quart jars). Using a different syringe for attempts 2 and 2.5 (5 jars each time ) blip of anything grew in jars. Starting over with third syringe and final syringe for 5 more jars it appears the growth is present but is monokaryotIc (which almost made me shit my pants when I read because I was getting I'm at my wits end trying to research and put forth as much effort as I have ). I honestly in my infinite glory of noobness think that second syringe (used for attempts 2 and 2.5) may have been bunk but I have zero friends/ input except for you guys to guide me in that determination An more importantly how to further refine my practice for my next attempt with the new batch which arrived sometime between tonight and Saturday ( had temps in 20s here last night but ive read conflicted views on this, seemingly in favor of they should be ok if they did freeze a little)
Again thanks for reading and helping out, I'm actually quite excited again
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