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Offlinejohnnytheswede
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PF Tek with agar instead of syringe?
    #20496401 - 08/30/14 05:14 AM (10 years, 4 months ago)

Hi Forum,

I am collecting materials and knowledge to start my first culture and I live in a country where syringes are *very* hard to come by. I am also inspired by agar which would enable me to select a strain which has more rizomorphic properties and my idea is to germinate spores on agar and select a petri dish which has good rizomorphic growth, and take the contents of this dish and put it inside a conventional jar of pre-sterilized vermiculite-rice flour substrate and then incubate it and continue with the normal PF Tek method.

From what I have learned, I understand that the transfers are the main entry points for contaminations. This means that when I open the jars with the substrate this needs to be done in an environment with very little mold spores and done quickly. To this end I have crafted a Glove Box which has latex gloves duct-taped to sleeves so that NO outside air can come in. Further, to purify the air inside I have installed a HEPA filter directly on the box and also attached a socket to the box so that I can run my vacuum cleaner removing all air from the box replacing all the air in the box with HEPA-filtered air. I will also have a small mister with bleach inside the box and alternate bleaching and air evacuation (using the attached vacuum cleaner) to ensure sterility. What do you think of this idea? I hope that it will enable me to safely transfer the culture from the petri dish to the PF Tek jar with less probability for contamination. I also hope that through this, I can be less strict about cleanliness outside the box itself since everything is done inside the box. No need to pause and pull out syringe outside the box and flame it, I'll have one pre-sterilized ordinary kitchen implement for every petri dish also removing the need for a flamer.

I am also a bit curious about something that seems like opposing pieces of advice. In the RR-video, we see that the PF Tek is described as involving the birthing of the cakes to require soaking of the cakes for 24 hours... will this not mean that the mycelium will drown? Of course not, they seem to know what they are doing, but please explain this a bit further, it seems risky because there is another video on Youtube, which I believe you know of (about one hour long, called "Mushroom Growing Made Easy") in which they say that "Do not touch the mycelium", "Mycelium will be harmed if water drops on to it", etc. Watching that video made me think, "Oh I will not harm you poor little mycelium", and then watching the RR video I thought "Oh no! Be careful! What are you doing!?". I would be very grateful for any clarification here.

Kind regards
Johnny


--------------------
Just learning to cultivate mushrooms, immensely impressed by Paul Stamets ("Pair up with Fungi!") and the knowledge and helpfulness at The Shroomery. Concentrating on Gourmet and Medicinal Mushrooms: Reishi, Shiitake, Garden Giant, Lion's Mane, Turkey Tail.

Acronyms (for noobs like me): http://www.shroomery.org/forums/showflat.php/Number/5875173#Post5875173
Have opted out of rating system because of personal reasons, read my bio if you are interested in why. (I opted out before receiving or giving any ratings, meaning no offense to anyone.)

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OfflineNos-hroom
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Re: PF Tek with agar instead of syringe? [Re: johnnytheswede]
    #20496413 - 08/30/14 05:29 AM (10 years, 4 months ago)

So far, I can recommend detaching the gloves from the glovebox, and dunking or soaking cakes for 24 hours is no big threat. They'll survive that. However, don't forget about them. The organism will die after 48 hours, and 24 hours is plenty as far as I'd advise.

As for not touching mycelium, the stuff is usually tough as nails (in the face of contaminants) once it has fully colonized the substrate, though it certainly will bruise if you don't handle it gently and properly.

I can't honestly give input on whether agar would or would not do well with BRF from experience, but it certainly could, hypothetically, have a slower rate of colonization due to the dense substrate in question and the more substantial properties of agar. I'd wait for a more experienced BRF user to chime in, here.

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OfflineFivel
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Re: PF Tek with agar instead of syringe? [Re: Nos-hroom]
    #20496504 - 08/30/14 06:24 AM (10 years, 4 months ago)

I have use agar to inoculate Pf tek cakes. I would advise you to make a still air box instead of the glove box, and I would also not skip the flame sterilization of your scalpel. Nothing beats flame sterilization IMO. What I did was cut one plate into 10 wedges and used 2 wedges per jar, just sat directly on top of the cake inside the jar. I used filtered gas exchange ie. poly stuffed lids, and i skipped the dry verm layer. It worked really well.


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

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Invisiblemycomattie
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Re: PF Tek with agar instead of syringe? [Re: Fivel]
    #20496780 - 08/30/14 08:54 AM (10 years, 4 months ago)

Besides the inability to shake (one of the best advantages of using grains, WBS, etc.), how would you get past the vermiculite layer on PF style jars with the agar wedges?


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

Edited by mycomattie (08/30/14 08:55 AM)

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OfflineFivel
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Re: PF Tek with agar instead of syringe? [Re: mycomattie]
    #20496789 - 08/30/14 08:57 AM (10 years, 4 months ago)

Quote:

mycomattie said:
Besides the inability to shake (one of the best advantages of using grains, WBS, etc.), how would you get past the vermiculite layer on PF style jars with the agar wedges?




I skipped the dry verm layer and use polyfill stuffed holes for GE on my lids.


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

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Offlinemushpunx
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Re: PF Tek with agar instead of syringe? [Re: mycomattie]
    #20496811 - 08/30/14 09:04 AM (10 years, 4 months ago)

Yea friend it was a cool idea but simpler is better in this case. Short of workinh in front of a flow hood (which uses hepa too) a Still Air Box is the best way to go.

Its just a clear plastic box with two arm holes cut in. The idea is to keep tge air still, and by using deliberate, careful motions (such as not moving anything over top of an open dish or jar), the contaminants wont settle into your sterile mediums. You can pull your arms in and out to flame sterilize and if I was to drop agar into a jar it is what I would use.


I dont know which other videos you watched, but the RR video is basically the gold standard. I just watched them recently and it sums up all the consistent information I had to search for and read.

Im honestly not sure if you can drop agar into a PF jar, I know you would have to skip the Dry vermiculite layer and come up with a better lid.

If the only thing holding you back is a syringe Im sure there is another way to get one. I mean, you can get spores and hepa filters.. and you have the internet so why not just order one? It doesnt have to have spores in it. If you have a spore print and a syringe (even one used for printer ink) that you can sterilize then you can make a spore syringe yourself.

If you have a pressure Cooker, you might as well do grains instead of PF tek if you are going to put the time and energy into agar.
But if you dont have one, I would buy one before messing around with agar to BRF jars and in the meantime use a syringe.

Im a herion addict so I know you can get or make a syringe anywhere!


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

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Edited by mushpunx (08/30/14 09:14 AM)

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Offlinekmetric
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Re: PF Tek with agar instead of syringe? [Re: Fivel]
    #20496817 - 08/30/14 09:09 AM (10 years, 4 months ago)

Agar to cakes would work, like Fivel said you just skip the verm layer and make sure you have some sort of GE going on. I know some experienced cultivators use cakes as a quicker way of testing out their isolates to see which ones they should go to bulk with.

You seem to have done your homework so far, and if you're going the agar route already, have you thought about going agar->grain? You can make a master grain jar from agar, and use that to make even more jars. Either way good luck :cool:

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OfflineFivel
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Re: PF Tek with agar instead of syringe? [Re: kmetric]
    #20496850 - 08/30/14 09:19 AM (10 years, 4 months ago)

I'm telling you that I have already done agar to cakes. It worked great. It was awesome watching an isolate annihilate that cake. Like a billion fingered hand came down and just engulfed it. Or like upside down white flames. :rockon: It works. It took my cakes 24 days to fully colonize. 1/2 pint wide mouths.


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

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Offlinemushpunx
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Re: PF Tek with agar instead of syringe? [Re: Fivel]
    #20496875 - 08/30/14 09:29 AM (10 years, 4 months ago)

^ Thay sounds so fucking cool! Blowing up some cakes!

So you  just skip the verm layer, drop the wedge on top the BRF inside a still box and put a filter lid on?


Ive never had a cake go bad from traditional PF tek though. Unless you are testing something out I would either do grains if I had a pressure cooker or find some way to get a needle if I was in the OPs situation


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

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AMU Q&A

Edited by mushpunx (08/30/14 09:30 AM)

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OfflineFivel
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Re: PF Tek with agar instead of syringe? [Re: mushpunx]
    #20496894 - 08/30/14 09:39 AM (10 years, 4 months ago)

Well I mean if he wants to use agar to go to cakes I say go for it if you are confident in your skills. I used agar my first grow cause I had to to save everything. Long story but all my shit got fucked and I was left with contaminated LC's that I had to put to agar to save since I didn't have the money to get another syringe. I transferred out 5 or 6 times before I started seeing sectoring. When I noced those pf jars I used 1 plate to 5 jars. I cut one plate into 10 wedges and used 2 wedges per jar, flaming the scalpel in between each jar. In a still air box. Everything wiped down with alcohol and I have a bleach soaked towel on the bottom for the 'floor' of my SAB. Quick motions. One wedge at a time replacing the lid on every jar after each motion. 2 wedges at a time / flame / 2 more on another jar / flame / I didn't pay attention to where the wedges landed I just threw them in there as quickly and smoothly as possible. I didn't get one contam. I only did 10 jars though. Sorry to be a little off topic and not to hijack your thread I just wanted to say that if you are smart you can start off with agar.

It just sounds like OP isn't tryin to get stoned with his friends. He seems to have passion. I bet he succeeds too because of it. :thumbup:


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

Edited by Fivel (08/30/14 09:41 AM)

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Offlinemushpunx
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Re: PF Tek with agar instead of syringe? [Re: Fivel]
    #20496923 - 08/30/14 09:59 AM (10 years, 4 months ago)

Well F ivel has me convinced. Go for it!!

Read about sterile technique and watch RR in front of a flow hood. Same motions just in a still air box instead.

Can you use Agar without a pressure cooker?


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

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OfflineNos-hroom
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Re: PF Tek with agar instead of syringe? [Re: mushpunx]
    #20496948 - 08/30/14 10:14 AM (10 years, 4 months ago)

In theory, you could buy "sterile" pre-made plates and use syringes made by people with more sterile conditions. However, I'd say getting the PC would unlock the gate to dozens of at-home projects, let alone agar.

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OfflineFivel
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Re: PF Tek with agar instead of syringe? [Re: mushpunx]
    #20496949 - 08/30/14 10:14 AM (10 years, 4 months ago)

:whathesaid:

Quote:

mushpunx said:
Can you use Agar without a pressure cooker?





This is going to be the hard part IMO. I use no pour so I pressure cook mine. I don't know if you can pour inside of a SAB or not. I would look into the agar first and make sure you can accomplish making clean plates OP. What tek(s), if any, were you planning on using? Do you have a pressure cooker?


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

Edited by Fivel (08/30/14 10:14 AM)

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OfflineBurberrySpores
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Re: PF Tek with agar instead of syringe? [Re: Fivel]
    #20497016 - 08/30/14 10:39 AM (10 years, 4 months ago)

Op.

You need to go get a Pc to step up your shroomery game.

If you are working with agar

You should start doing grains IMO :thumbup:


--------------------
I could go crazy... and no one would notice :seeya:

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Offlinejohnnytheswede
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Re: PF Tek with agar instead of syringe? [Re: BurberrySpores]
    #20500544 - 08/31/14 04:01 AM (10 years, 4 months ago)

Thank you for your overwhelming responses :-)

My knives *would* be flame sterilized but that would happen *before* I put them and everything else in the box, I would keep them immersed in a bleach bath (inside the box) after flame sterilization avoiding having to handle anything outside the box. I was a bit unclear, what I meant is that I would not have to flame sterilize *during transfer procedures*, just pick one of my 10 pre-sterilized implements and use it once and then put it aside. This would also enable me to work in not only still air, but almost totally clean air. Thank you though for the explanation of how a Still Air Box works.

Further, I will await full colonisation of the jars.

I will try to do PF Tek with agar in the way described by this thread but as a backup I also ordered syringes off the net, we'll see how picky customs are, you know they sometimes x-ray stuff and if they see a syringe, then bye bye! (To keep a low profile I ordered 2 syringes with oyster mushroom spores.)

Some further questions, the meaning of some acronyms:

GE

BRF

WBS

LC

I do know what a PC is and I've got one! I got some teflon hose too so I will use it to distill water.

From the context of people's replies I guess OP is me, but what does the acronym stand for?

BTW: I tried to check the list of terms and unfortunately it does not work (404)... who do I ask to fix it?

Kind regards
Johnny


--------------------
Just learning to cultivate mushrooms, immensely impressed by Paul Stamets ("Pair up with Fungi!") and the knowledge and helpfulness at The Shroomery. Concentrating on Gourmet and Medicinal Mushrooms: Reishi, Shiitake, Garden Giant, Lion's Mane, Turkey Tail.

Acronyms (for noobs like me): http://www.shroomery.org/forums/showflat.php/Number/5875173#Post5875173
Have opted out of rating system because of personal reasons, read my bio if you are interested in why. (I opted out before receiving or giving any ratings, meaning no offense to anyone.)

Edited by johnnytheswede (01/04/15 08:37 AM)

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OfflinePsiloBeast
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Re: PF Tek with agar instead of syringe? [Re: johnnytheswede]
    #20500566 - 08/31/14 04:15 AM (10 years, 4 months ago)

Shit, I take mushrooms to get fuckered up

See shit, ya know?  :laugh2:

:goodluck:


--------------------
Whosoever is delighted in solitude is either a wild animal or a god...
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Water Extraction Tek,Frank's Monotub Tek ,Do What He Does...

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OfflineFivel
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Re: PF Tek with agar instead of syringe? [Re: PsiloBeast]
    #20501009 - 08/31/14 09:28 AM (10 years, 4 months ago)

Well we all suggest a still air box or SAB because it works better than a glove box. It's cleaner than a glovebox. These are time tested methods by people who do this for a living. That's why we suggest them. As for your acronyms they are:

GE - Gas Exchange

BRF - Brown Rice Flour

WBS - Wild Bird Seed

Lc - Liquid Culture

Having a pressure cooker helps greatly. Also, you won't be using distilled water unless you are making syringes yourself. Distilled water isn't used during the fruiting stage or in the cake mix / substrate because it will leach nutrients from your mushies. Use tap water. As for flame sterilizing your utensils and setting them in a bleach bath I am just going to go on a limb and say that won't work as well as just using one scalpel and flame sterilizing it between uses. Bleaching something will sanitize it, but not sterilize it. You can look this up for yourself if you don't believe me. This is why we use SAB's as well because it makes it safer to flame sterilize. Glove boxes don't work as well because when you move your hands you are creating currents from the vacuum seal of the gloves to the box. If that makes sense. The more you read on here the more you realize that the reason people do the things the way they do them is because they have tried everything else already and we are left with the stuff that works.. simply put.


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

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Offlinemushpunx
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Re: PF Tek with agar instead of syringe? [Re: Fivel]
    #20501171 - 08/31/14 10:10 AM (10 years, 4 months ago)

! Oh if you have a pressure cooker you're set my friend, you could drop those agar wedges right into some jars of prepped grains.

Im not trying to discourage you from your agar to PF tek ambitions, but grain is much more versatile and very simple.

Grain allows you to inncoculate ten more quart jars from one quart of colonized grain

Also allows you to spawn to a bulk substrate for very big yeilds. Honestly I think grain is alot less work than PF tek. Most people start with cakes because they dont have a pressure cooker and heard its easier.


As for a syringe, if they dont show, you could sterilize a syringe meant for somethimg else (cooking, printer ink, epoxy glue etc) in your pressure cooker and make your spore syringe with that.

Good luck!


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

Amateur Mycologists United
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OfflineFivel
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Re: PF Tek with agar instead of syringe? [Re: mushpunx]
    #20501348 - 08/31/14 11:06 AM (10 years, 4 months ago)

Yeah man if you have a pressure cooker and you want to just go grains, I would think you can no problem. This tek will get you going man it did me. I got mushrooms after I did this.


--------------------
"The only way to deal with an unfree world is to become so absolutely free that your very EXISTENCE is an act of rebellion."
- Albert Camus  :bonghit:

Have Orissa Prints for trade!

In search of PF albino, PF redspore, PF classic :pm:

"... the associations between sadism and GAIT (Global Assessment of Internet Trolling) scores were so strong that it might be said that online trolls are prototypical everyday sadists."

:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2::meltdown::mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

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Offlinejohnnytheswede
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Re: PF Tek with agar instead of syringe? [Re: Fivel]
    #20502077 - 08/31/14 02:40 PM (10 years, 4 months ago)

Thank you everyone for your advice and sharing your knowledge! I'll tell you more in a couple of months when I have my own real experiences.

Best regards
Johnny


--------------------
Just learning to cultivate mushrooms, immensely impressed by Paul Stamets ("Pair up with Fungi!") and the knowledge and helpfulness at The Shroomery. Concentrating on Gourmet and Medicinal Mushrooms: Reishi, Shiitake, Garden Giant, Lion's Mane, Turkey Tail.

Acronyms (for noobs like me): http://www.shroomery.org/forums/showflat.php/Number/5875173#Post5875173
Have opted out of rating system because of personal reasons, read my bio if you are interested in why. (I opted out before receiving or giving any ratings, meaning no offense to anyone.)

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