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Offlinemasarap
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Some questions after first attempt and failure
    #8682568 - 07/26/08 12:06 AM (16 years, 5 months ago)

This was my first attempt to grow.  I inoculated 4 1/2 pint jars.  Today is the 4th day after inoculation.  All jars have the green mold. 

I followed the pf tek by MushroomsRR
http://www.youtube.com/watch?v=ZHJQrsZFQdE


Except I used a pressure cooker (for the first time).  I think this is where I fatally messed up.  I have a cheap pressure cooker w/o a pressure gage.  The cooker I have is a 'rocker' by presto.  I never saw the knob thing rock like its meant to.  After the cooker  locked I assumed it was all good and maybe the rocker was broken and started timing the cook.  But looking at the rocker now I noticed its adjustable and it was turned to the tightest setting (basically no steam was escaping) and almost all the watered remained on the bottom after 50mins(another red flag).  I was rushed and even though I had that feeling I messed up I continued w/ the inoculation. 

Well thats my n00b story.

My questions for my next attempt..


1.  The syringe I used still has like 4cc left in it, should I completely discard this or will it be safe to use again? - I have another fresh never opened syringe so Im not desperate.

2.  Can I or is it recommended I spray my inoculation chamber w/ Lysol prior to inoculation?

3.  The tape over the holes on jars.  Which tape works best?

Edited by masarap (07/26/08 12:12 AM)

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OfflineSkeeblix
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Re: Some questions after first attempt and failure [Re: masarap]
    #8682604 - 07/26/08 12:21 AM (16 years, 5 months ago)

What's the problem here? It's perfectly fine for there to still be plenty of water in your PC when you're done.

If you're saying no steam was escaping the rocker, it probably did not pressurize enough to make steam escape. I don't know the weight settings on your particular model, but so long as it hit 10 PSI or over during the cook, your jars should be perfectly fine.

Don't count yourself out right away because you don't know if you messed up or not. Nothing you did leads to any indications of  improper procedure.

Keep in mind, you didn't say much about how you went about your inoculation, but don't call it a failure yet.

You'd be surprised how resilient mushroom mycelium can be, and the kind of conditions is will still show up in.

I hope this turns out nicely to shake your newbie paranoia off and get you going with a can-do attitude. Nothing in this hobby is very hard, it just has to be done right, and done clean. If you can follow a tek and at least halfway keep things sterile, then provide the proper environment, the mushrooms themselves do all the REAL work.

As for your questions:

1.You can use every last bit of spore solution in a syringe, provided you haven't gone around sucking things up into the syringe to contaminate it.

2. I believe you're talking about a glovebox, a still air workspace to do inoculations or other particularly contaminate-risky procedures, like grain to grain transfers and the like. Ideally, yes, you do spray out the inside of your work area with some kind of disinfectant, or at least some alcohol solution. Take note to let the fumes dissipate a bit inside the box before you use open flame to sterilize your needle inside the glovebox.

3. 3M micropore tape is good stuff for covering jar holes if you're really that paranoid about leaving them open. You can find it in the medicine aisle of just about any little store, like Walgreens and shit.


--------------------
This post approved by:


Premedman1 said:
:lol: I just shat my pants.

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InvisibleWolfgang
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Re: Some questions after first attempt and failure [Re: masarap]
    #8682608 - 07/26/08 12:23 AM (16 years, 5 months ago)

1. Save it and try again.. it may or may not be the syringes fault.

2. I wipe it down with 76% alcohol (which is far better and safer than lysol)

3. The pf tek doesn't require any tape over the jars that's why we put the dry verm layer on top..  it acts as a contam barrier and a filter just like micropore tape would

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Offlinemasarap
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Re: Some questions after first attempt and failure [Re: masarap]
    #8682625 - 07/26/08 12:31 AM (16 years, 5 months ago)

Oh yes.  My inoculation.  Wheww.. I also did something really stupid there.  I did it without a glove box or chamber.. In a somewhat humid basement.  Also I used a candle to heat the needle :blush: b/c the lighter I had wasn't working to great. 

I used gloves, alcohol and a candle for the needle.

Besides not using a glove box I believe the candle was a poor choice as well.  Yes the needle got red hot but there was a black residue from the candle (maybe collecting mold spores after cooling)

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OfflineBrennus
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Re: Some questions after first attempt and failure [Re: Wolfgang]
    #8682630 - 07/26/08 12:34 AM (16 years, 5 months ago)

If you research liquid culture teks, primarily with honey water, you can turn that 4cc's from your first spore syringe (which probably isn't really enough to knock up your next batch of jars) into several dozen syringes of mycellium that will colonize faster. Here's a link to the one I used: http://www.shroomery.org/forums/showflat.php/Cat/0/Number/5874305/page/0/fpart/all/vc/1

(I used live spores instead of a cutting and it appears to have worked)

If you're that concerned with the pressure cooker, the PF Tek works fine with a regular pot and steam. It takes a little more than 90 minutes, but it's worth it if your PC is sketchy. Also, tape doesn't really matter for the PF Tek - the dry vermiculite barrier is there to prevent contams from entering the jars via the holes in the jar.

The black marks from the candle flame are soot. When I inoculate, I hold the needle over a candle flame for 10 seconds or so, then wipe it with alcohol before going back into my glove box. If you got contams from the inoculation point and not from improperly sterilized jars, it's more likely because you weren't using a glove box.

One last thing - buy yourself a cheap rubbermaid from Walmart and make a glove box. Best $8 and 45 minutes I ever spent for this hobby.

Edited by Brennus (07/26/08 12:37 AM)

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Offlinemasarap
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Re: Some questions after first attempt and failure [Re: masarap]
    #8682634 - 07/26/08 12:35 AM (16 years, 5 months ago)

Thank you for your replies.  I know I completely messed this one up and will be much more careful next time.

Now I'm confused about the tape b/c in the pf tek video he uses micropore tape?  So its really not necessary?

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OfflineBrennus
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Re: Some questions after first attempt and failure [Re: masarap]
    #8682644 - 07/26/08 12:38 AM (16 years, 5 months ago)

RR himself has said tape isn't necessary for the PF Tek on the forums numerous times due to the dry vermiculite barrier.

Micropore tape can also be substituted for regular ol' medical tape from your first aid kit, too.

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Offlinemasarap
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Re: Some questions after first attempt and failure [Re: masarap]
    #8682650 - 07/26/08 12:42 AM (16 years, 5 months ago)

Thank you.  Alright I already feel better about starting a new batch.  But first and foremost I will experiment w/ my cooker and see If i can get it to 'rock' like it is said to do.  And yes I will use a glove box next time for sure.

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Offlinemasarap
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Re: Some questions after first attempt and failure [Re: Brennus]
    #8682669 - 07/26/08 12:54 AM (16 years, 5 months ago)

Quote:

Brennus said:
When I inoculate, I hold the needle over a candle flame for 10 seconds or so, then wipe it with alcohol before going back into my glove box.




Thats the other thing..I wasn't whipping my needle with alcohol either.


Thank you for all your help.  I'm going to make some new cakes tonight assuming I can get the cooker to "rock" :laugh:

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OfflinePowerOfTheCoir
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Re: Some questions after first attempt and failure [Re: masarap]
    #8682700 - 07/26/08 01:02 AM (16 years, 5 months ago)

Your pressure cooker will definitely let steam out when it gets up to pressure - otherwise it would have exploded. Probably not the problem though - boiling water with no extra pressure works for pf cakes.

If you use take, use micropore or transpore. It must be breathable. But I agree with everyone else, you don't need any tape at all.

Did you notice the green funk appear at the inoculation points, or in areas not associated with the inoculation points? That will give you a good idea of what stage caused your problems.

Notice how foil is often put over the jar tops during sterilization. Keep this on there until right before you inoculate. It keeps contaminants from falling into the injection points and riding the needle down into the substrate. Once you've injected, you can throw the foil away.


--------------------
Check out my first ever TEK!
Shroom capsules with the Cap-M-Quik (pics)

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Offlinemasarap
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Re: Some questions after first attempt and failure [Re: masarap]
    #8682714 - 07/26/08 01:12 AM (16 years, 5 months ago)

As far as i can tell the green mold is growing only on the white mycelium.  All the white has some green.  Theres no green growing else ware so yes I guess the inoculation was the kill?

** also yes the foil.  I had taken it off first to check the tape, I was curious If it had melted.  Then took off all the foil at least a good few mins before I inoculated.

And as far as the tape I used masking tape which I'm guessing wasn't that great.

Edited by masarap (07/26/08 01:21 AM)

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OfflineRogerRabbitM
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Re: Some questions after first attempt and failure [Re: masarap]
    #8683443 - 07/26/08 09:02 AM (16 years, 5 months ago)

If you use masking tape, remove it after inoculation.  If you use micropore tape, you can leave it in place.  Tape isn't 'necessary' but it's an extra precaution.

Soot on the needle from your candle doesn't hurt. I advise against wiping the needles with anything.  You can't get cleaner than 'red-hot'.

If the whole jars are green, your sterilization process didn't work. If the green is only at the inoculation sites, then your inoculation procedure or syringe itself is the problem. Use a simple kettle to boil as per the video, or use a pressure cooker properly.

Consider a glovebox a must.  It isn't necessary to use lysol or anything else in a glovebox.  Gloveboxes are only a means of stopping air currents in the room, nothing else.

I'd discard the used syringe and use your new one.  Since you didn't use a glovebox, there's a chance that room air got sucked into the syringe, so why take a chance?

The same videos that are on youtube are also on my website, but in much higher resolution, provided you have high speed internet service.  There's also text that fills in some of the questions that can't be easily covered in video format.  Read the brf tek page and the terrarium tek page.  The link is in my signature.
RR


--------------------
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"I've never had a failed experiment.  I've only discovered 10,000 methods which do not work."
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Offlinemasarap
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Re: Some questions after first attempt and failure [Re: RogerRabbit]
    #8684488 - 07/26/08 02:37 PM (16 years, 5 months ago)

Thank you for all the input.


I just finished my new inoculation of 5 fresh jars. I used the 'oven tek' for my inoculation site.

Captains Oven Tek

This time I was much more careful.


Looking at my old jars (now on day 5) the green mold seems to be all over, not just the inoculation sites.  I'm guessing when I used the pressure cooker the first time it never fully sterilized.  Last night the "rocker" actually rocked like it is meant to.  I feel better now knowing that.


So my new jars..

finished cooking at 4:30am last night

cooker was put aside to cool overnight

Around 2:30pm (about 10hrs later) today the cooker was opened, jars removed and inoculated immediately there after.

Jars are now in ventilated covered box at 84.8 F and 40%rH.  The temp goes down at night around mid 70s and peaks 87 at max.

I will update what happens in a few days.



Thanks again for all the great input  :thumbup:

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Offlinemasarap
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Re: Some questions after first attempt and failure [Re: masarap]
    #8705426 - 07/31/08 12:23 AM (16 years, 5 months ago)


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Offlinemasarap
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Re: Some questions after first attempt and failure [Re: masarap]
    #8705735 - 07/31/08 01:42 AM (16 years, 5 months ago)

I conclude to anyone out there planning to grow for the first time...
Do NOT! take sterility for granted.
Do NOT! be in a Rush.  Plan carefully how you will do things. 

On my new jars I used the Oven Tek.  Captains-Oven-Tek

What I did...
-Preheat oven 200F
-Unwrapping the aluminum foil on jars only when the jar was already on the oven rack (I pulled out the rack only enough so the door wouldn't burn me while I'm working... I guess as long as you have that hot air coming up your good)
-Wipe the top of jar with alcohol
-Flame sterilize needle
-Immediately after needle is red hot inoculate jar (all four holes).
-Take jar off rack and set aside
Repeat process for next jar.

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InvisibleKrez
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Re: Some questions after first attempt and failure [Re: masarap]
    #8705740 - 07/31/08 01:44 AM (16 years, 5 months ago)

Ditch the oven tek and build yourself a glovebox


--------------------
Kids, you tried your best and you failed miserably. The lesson is, never try.  (Homer)

Look, the thing about my family is there's five of us. Marge, Bart, Girl Bart, the one who doesn't talk, and the fat guy. How I loathe him. (Drunk Homer)

All the ways you wish you could be, that's me. I look like you wanna look, I fuck like you wanna fuck, I am smart, capable, and most importantly, I am free in all the ways that you are not. (Tyler Durden)

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OfflineSelfDestruct
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Re: Some questions after first attempt and failure [Re: Krez]
    #8705762 - 07/31/08 01:54 AM (16 years, 5 months ago)

Yeah from what i've read numerous times around here, oven tek is pretty bad.  If you're gonna do open-air inoc make sure you do it in a very clean place.  Thankfully my open-airs have been contam-free thus far *knocks on wood*

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Offlinemasarap
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Re: Some questions after first attempt and failure [Re: Krez]
    #8705767 - 07/31/08 01:55 AM (16 years, 5 months ago)

By all means I'm not challenging the glovebox.  This is only my personal experience.

And yes the glovebox seems much safer.  Agreed.



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InvisibleKrez
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Re: Some questions after first attempt and failure [Re: SelfDestruct]
    #8705769 - 07/31/08 01:56 AM (16 years, 5 months ago)

Yea just shut the ac off and any fans...Oust the shit out of the room and remember to flame the syringe.


--------------------
Kids, you tried your best and you failed miserably. The lesson is, never try.  (Homer)

Look, the thing about my family is there's five of us. Marge, Bart, Girl Bart, the one who doesn't talk, and the fat guy. How I loathe him. (Drunk Homer)

All the ways you wish you could be, that's me. I look like you wanna look, I fuck like you wanna fuck, I am smart, capable, and most importantly, I am free in all the ways that you are not. (Tyler Durden)

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Offlinerisndeath
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Re: Some questions after first attempt and failure [Re: Krez]
    #8705848 - 07/31/08 02:37 AM (16 years, 5 months ago)

for precation i douse my latex gloves before inoculating which is the time you are most likely to introduce contams to the substrate, i use 91% alchahol on the foil where the 4 holes are to make sure i catch any particles near the inoculation holes. flame the needle and let it sit and cool for a little inside a towel that has been doused with alchahol.

i also use a ton-load of lysol on everything i spray myself, the air in the work inviroment, the jars, all counter tops, and then I inoculate the jars.

they say it is ok once the jar has been inoculated to remove any barrier like "foil/lid" from the jar because it will only restrict air and since you have a 1/2 to 1 inch of dry verm on the top layer this catches all contam worthy particles from entering the jar and contaminating your substrate, so thats what I do.

I made sure I was super sterile since I read alot of people have contams, so far I have none *keeps fingers crossed*


--------------------
I am the fakest person on this website all claims made by this account are entierly fictional, i am a loser in real life and i pretend to know everything on this website as a way to increase my self esteem in no way am i contributing to the use of mushrooms or anything said on this forum.

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