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OfflineYaMoonSun
The Double Standard


Registered: 10/23/14
Posts: 3,967
Loc: NY
Last seen: 5 months, 10 days
Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
    #22450020 - 10/29/15 04:58 PM (8 years, 3 months ago)

Watch my cup of noodles be the only one that doesn't contaminate :lol:


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Offlinemushpunx
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Registered: 04/20/14
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
    #22450156 - 10/29/15 05:26 PM (8 years, 3 months ago)

Are you printing in your still air box?

You want to prep the SAB by spraying the walls lightly with soapy water. Wear freshly laundered clothes, tyvek sleeves if you have them, gloves, scrub arms like a surgeon, face mask, wipe gloves down with ISO.
You need to know how to use an SAB to utilize it properly, you never move anything over top sterile work, use controlled desicisive hand motions to keep the air as still as possible.

For printing you need to tear off a piece to get to the clean foil. Cut the cap with a sterile scalpel, fork the top...  place onto foil, after 24 hrs remove cap , let dry in still air box, carefull place in fresh (fresh is sterile) ziplock. Most people fold in sterile foil envelope first.


That you looked up the syringe making on YouTube is not helpful. I asked you to describe your process.

We need to know that you are properly sterilizing your syringes, water, tools etc.

You need to sterilize a shot glass, wrapped up in foil so that it can be opened in your still air box. The syringe should be filled with water and sterilized wrapped in foil as well. A jar of water should also be sterilized and allowed to cool. Some people add a couple drops of sex lube in prior to sterilization.

You carefull open your print in still air box following the tips I gave you. You would flame sterilize your syringe and drop a couple drops onto the print where you intend to use, and with a flame sterilized loop loosen up that section, and pour off the print into the shot glass with however much sterile water in it. Flame the syringe and pull up the water and cap.




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

Amateur Mycologists United
AMU Q&A


Edited by mushpunx (10/29/15 05:39 PM)


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OfflineYaMoonSun
The Double Standard


Registered: 10/23/14
Posts: 3,967
Loc: NY
Last seen: 5 months, 10 days
Re: Contaminated, again; Six out of Six [Re: mushpunx]
    #22450253 - 10/29/15 05:46 PM (8 years, 3 months ago)

No, I didn't print in the SAB, the point of the glass was to prevent air flow :shrug:



My SAB is pretty whack :lol: It probably needs to be redesigned.
It's just a temporary fix to prevent contamination from getting into the sterile shot-glass. The syringe is already made :shrug:


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Offlinemushpunx
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
    #22450278 - 10/29/15 05:51 PM (8 years, 3 months ago)

Quote:

YaMoonSun said:
No, I didn't print in the SAB, the point of the glass was to prevent air flow :shrug:



My SAB is pretty whack :lol: It probably needs to be redesigned.
It's just a temporary fix to prevent contamination from getting into the sterile shot-glass. The syringe is already made :shrug:




Covering the print is fine, but what about all the contams landing on your foil before you even put the cap down? Printing in open air is only suitable for prints that are going to be cleaned up on agar.

Well Im almost certain your contams are coming from your syringes, friend.  Unless you walk us through how you are printing and syringe making step by step I doubt we can help you solve your problem


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

Amateur Mycologists United
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OfflineYaMoonSun
The Double Standard


Registered: 10/23/14
Posts: 3,967
Loc: NY
Last seen: 5 months, 10 days
Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
    #22450313 - 10/29/15 05:59 PM (8 years, 3 months ago)

Quote:

I asked you to describe your process.





Usually, with success

• Boil water for ten minutes to sterilize
• Draw sterile water into the syringe and expel into nearby glass eight times
• Draw sterile water into syringe once more, and store it in the fridge to cool
• Use 70% isopropyl alcohol to sanitize a shotglass using half a paper towel, drying with the other half
• Flame sterilize a blade, one that won't tear the aluminum
• Scrape a few spores into the shotglass
• Flame sterilize the syringe and eject the sterile water into the spores, and suck them back into the syringe quickly, but accurately, as not to get any bubbles.

The print making doesn't have but three steps... I already told you what they were. The SAB was made last night; Hopefully that solves all the problems :pipesmoke:


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OfflineMachiavelliavore
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Re: Contaminated, again; Six out of Six [Re: Zuul]
    #22450331 - 10/29/15 06:03 PM (8 years, 3 months ago)

Quote:

Zuul said:
Are you reusing the same jars over and over?  With all the different variables it's not going to be easy trying to pin point the cause of contamination. If these cakes don't make it toss everything, get a PC, and get fresh everything. There's a turd in your punch bowl and it's not going to be easy fishing it out.




...Sterilizing the substrate sterilizes the jar...

I agree with punx, it's probably your syringe-making that's the problem.  PF tek was designed for open air inoculation.  Are you using fine verm for the barrier?  If not, you might wanna invest in some silicon sealant and make injection ports and a tyvek covered GE hole.  It's only like $3 at ace, tyvek free at the post office.

Definitely sterilize whatever you scrape spores into.  The guide I read (off this site's tek section) said to whipe it down with alcohol.  Also, when you lower the syringe to suck up spores, tilt the container sideways so anything falling off your hands or the syringe falls on the side of the glass or the floor of the air box.  I'm surprised my first grow wasn't a flop.  Sounds like you read the same guide I did.  Whipe your syringe down as well, or just put it in a piece of foil in your steam sterilizer.


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


I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


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Offlinemushpunx
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
    #22450537 - 10/29/15 06:54 PM (8 years, 3 months ago)

YaMoonSun said:
Quote:

I asked you to describe your process.





Usually, with success

• Boil water for ten minutes to sterilize
• Draw sterile water into the syringe and expel into nearby glass eight times
• Draw sterile water into syringe once more, and store it in the fridge to cool
• Use 70% isopropyl alcohol to sanitize a shotglass using half a paper towel, drying with the other half
• Flame sterilize a blade, one that won't tear the aluminum
• Scrape a few spores into the shotglass
• Flame sterilize the syringe and eject the sterile water into the spores, and suck them back into the syringe quickly, but accurately, as not to get any bubbles.


Here's one of your main problems. You have the term "sanitize" correct.. you arent sterilizing the shot glass. If you dont believe me place a drop of ISO on an agar plate and watch what grows.
Even if you did sterilize it, touching it with a paper towel to dry it un sterilized it.

I believe that is where the bulk of your contamination is coming from. We usually wrap the shotglass up in foil and sterilize in a pressure cooker cycle, then only open the foil in still air box.

Another less serious issue is scraping spores with blade. It fucks up the spores. Loosen thrm with a few drops of sterile water (on print in still air box) scrape them up with a flame sterilized loop (easy to make) and pour the spores/drops of water into the sterile shot glass.


  Any lapse in sterile technique is enough to doom your syringe. Moving your gloves over top your open print or over your sterile shot glass can knock contaminants in. Even disturbing the air too much can move contams to settle into exposed sterile containers.

I dont know if there is a way to steam sterilize your shot glass PF style, or if you need to invest in a PC (which you most definitely should anyways), but if you correct these couple things it should go a long way.

I would toss any syringes you have already made dude





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

Amateur Mycologists United
AMU Q&A


Edited by mushpunx (10/29/15 07:05 PM)


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InvisibleSupalemonhaze
Spore syringe hater.
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Registered: 10/02/15
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Loc: 12" down Europe's butthole
Re: Contaminated, again; Six out of Six [Re: JacobStorm]
    #22450561 - 10/29/15 07:03 PM (8 years, 3 months ago)

Yea id definitely switch to brf if I didn't have a pc. One of the most forgiving substrate there is. Shouldn't have much trouble finding it. If I get it in my couuntry its a safe bet you'd find it anywhere else


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OfflineMadSeasonStudent
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Registered: 08/26/12
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Re: Contaminated, again; Six out of Six [Re: Supalemonhaze]
    #22450618 - 10/29/15 07:17 PM (8 years, 3 months ago)

You should of made up some agar or brf to test first.  I had a foul smelling cake that smelled like sour milk. I left in water for a week. I wouldn't fruit it.  Try leaving it longer on the stove next time.


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



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OfflineYaMoonSun
The Double Standard


Registered: 10/23/14
Posts: 3,967
Loc: NY
Last seen: 5 months, 10 days
Re: Contaminated, again; Six out of Six [Re: mushpunx]
    #22450677 - 10/29/15 07:30 PM (8 years, 3 months ago)

Bulk contamination came from sterilizing the shot glass wrong. Had never sterilized before and wanted to be extra causious that time around, and I goofed I guess. Had no prior issues with just sanitation. Also, yes, I swapped to brf, in regards to what haze said, and mad has a student? I'll be damned. I know dude, agar, I need it badly.


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OfflineMachiavelliavore
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Registered: 12/08/14
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Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
    #22450716 - 10/29/15 07:38 PM (8 years, 3 months ago)

Probably want to synch that foil down like a doo rag with some string.  Can you even do agar without a PC?  Maybe there are some microwave methods, I'm not sure.  I summon thee lord blackout to lay down your microwave wisdom on all us ignant bitches.

IME grains and cakes are about the same from spore syringe.  If your inoculant's good, your grow will be good.


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


I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


Edited by Machiavelliavore (10/29/15 07:39 PM)


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Offlinemushpunx
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Re: Contaminated, again; Six out of Six [Re: Machiavelliavore]
    #22450764 - 10/29/15 07:49 PM (8 years, 3 months ago)

A syringe is rarely clean enough for grains man. Its why we inoculate agar and make transfers u ntill the culture is clean enough to use as innoculant.
But BRF is a little more forgiving and will usually do fine from spore syringe.


OP I think you need a PC. Makes sterilizing a breeze. A presto 23qt is a great starter PC holss 10 quart jars at a time, theyre 60-70 bux.

Of course with a PC you could move onto grains and bulk  growing and agar work too!


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

Amateur Mycologists United
AMU Q&A


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OfflineMachiavelliavore
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Registered: 12/08/14
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Re: Contaminated, again; Six out of Six [Re: mushpunx]
    #22450861 - 10/29/15 08:05 PM (8 years, 3 months ago)

I've found grains to be more forgiving than cakes.  Probably cause I was using rye cakes which are a bit shitty in terms of texture IMO.

Mycellium seems to be very happy in grains, plenty of space to grow through, not sticky.


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


I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister.  I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave.  I think it will be good cause B+ is so potent.
Triggered yet?

Only a square would say "a cube is a cube."


No, this does not look right...


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OfflineYaMoonSun
The Double Standard


Registered: 10/23/14
Posts: 3,967
Loc: NY
Last seen: 5 months, 10 days
Re: Contaminated, again; Six out of Six [Re: Machiavelliavore]
    #22450940 - 10/29/15 08:18 PM (8 years, 3 months ago)

Quote:

microwave methods



:lol: You're right, I need that damn pressure cooker first. There's one jar outside that seems to not smell, but it hasn't shown signs of myc yet either

So it's agreed, OP needs a fucking PC
:nodofunderstanding:


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InvisibleJuiceh
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Registered: 09/25/12
Posts: 3,208
Re: Contaminated, again; Six out of Six [Re: YaMoonSun]
    #22451348 - 10/29/15 09:34 PM (8 years, 3 months ago)

Quote:

YaMoonSun said:
The syringe making I looked up on a YouTube video



Post the link to this. I've seen alot of terrible syringe tek videos on youtube that have huge flaws.


--------------------
Trade List

How I make 15 lbs of casing in a 941. How I line 12 shoeboxes with 1 39gal bag. How I Deep Fry Cubes!
RogerRabbit said:You need a bigger pressure cooker for this hobby. RR


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