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OfflineCamtaro420
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Cam's Grow #2. From Scratch. (Act:I.5 BRF Cake Phase)
    #19277624 - 12/15/13 03:26 AM (10 years, 2 months ago)

Hey Shroomery. It's me again.

So this time I'm going to just go with a grow journey here in this thread. My last one included some of the methods I was going to use to build my SAB, places I went and some of the supplies I ended up buying.

I wound up rushing into it a little bit and I used a really old syringe so nothing good but knowledge came from that experience. You can read all about it here.

Here's the new plan of my grow outline. I'll start by create some PDYA petris. Isolate a few good strong agar cultures. Scalpel out the best growth from the isolates to a few mother grain jars. Transfer grains from the mother jars to many more grain jars and eventually case and place in a mono.

I'm thinking smaller monos, nicely staked in my back closet with a GH. I want to do a grow similar to what Notahacker has listed in his "tale of 10 isolates" his work is solid and easy to follow so thanks for that. Got a whole walk in closet to put these bad boys in.

Wild bird seed for the grains. Maybe PP5 plastic for all the grain jars? My petris are all half pint's for the moment till the new ones come in from ebay(Updated 12/22 they finally came so my jar's will go back to being used for cakes).

Here's the equipment I am working with so far.

A SAB


My Sterilizer. I dunno if you guys saw the 25$ Steam pot I bought. Well f*ck that piece of sh*t. I'm cruising in style now.

Mother in law brought by the old All American Pressure Sterilizer from her old shroom days and I have taken up her torch.

It needed a new knob and a new gauge. Picked up the parts at my local hardware store and it worked great. (used a presto pressure gauge but it worked perfectly as it was the exact same design model of pressure gauge from AA.)When it got to 10 psi I had to weigh down the valve though. It's pretty old school.

Once I did it worked perfect got it to 15psi, maintained it for 30 mins. Viola the jar of liquid PDYA inside is done.

Update 1/6/14*
So nothing good came from the petris. I blame that mainly on the fact that a vendor syringe is not that reliable for agar work like other sources. Still a good read and the rest of my syringe that I used for BRF cakes seems to be going well.

I used a recipe found here. I used some substitutions though.

Potato Dextrose (Yeast-extract) Agar (PD(Y)A)
500ml water
5 g of instant potato flakes
9 g agar agar
10 ml corn syrup
1 gram nutritional yeast ('seasoning')

I used a coffee grinder that I use for dry shrooms and not coffee to grind all of the dry ingredients together (potato flakes, agar agar, yeast, till it was a fine powder. Then I boiled up the water added in the honey and corn syrup, made sure it was dissolved, then whisked the powdered ingredients in slowly but vigorously till it was all cooked in.

Afterward, I added this liquid strained through a small mesh strainer into another jar with the lid turned upside down so it would be easier to open and pressure cooked it at 15 psi for 30 minutes. Once it was cool enough to handle I added it and the jars to a SAB for pouring. The jars were cleaned with soap and water then dried with paper towels so they were not 100% sterile, but it's all I have till the mail gets here in a few days with the pre-sterile dishes.

I didn't use all the jars but most of them. The agar once poured in and closed up looked perfect inside but since they are not completely sterile. Back into the PC everything goes.

I have them all stacked up and cooking at 15 psi right now. Gonna blaze a fat bowl as the cooker works it's magic for the next 90 minutes and then let it cool till it's time to innoc in the morning.

On another note I also received these pre-made and sterilized petris[/url] from ebay seaweed solutions place. Going to use these tomorrow as well, less anyone has objections.


Here's the syringe. A+ Albino from a popular sponsored vendor.

Going to try to use as little as possible for each petri. I have about 15 so I will need to use less then 1 CC per jar/petri, but there is just a massive clumping of spores in this syringe I don't see how it wouldn't be enough for all these dishes.

Update 1/6/14 *
Not sure if I didn't use enough, or it was the contents inside, but nothing good came out of the syringe when added to the petris as mentioned above. I did however, use evilmusroom's take at the PF tek using the PC. So far so good results.

Update 12/22
I am going to innoc my BRF jars here in about 20-30 minutes. Today is gonna be a good day. Think I will also pour some of those agar dishes too.

Update 12/30
Ok so here is the news so far.

My PDYA jars grew nothing in the ways of fungus or molds. From what I can tell a handful of them grew some bacterial looking stuff, while the rest look like they did when I first PC'd them,

My 2/5 of my peptone beef agar petris wound up growing something that resembles cobweb mold, while the other three have nothing at all on them. Still trying to confirm wither or not if it's cobweb mold.

so all the agar I used is just wasted time and effort.

That's the bad news. 

The "could be" good news is, one of the four BRF cakes I inoculated 8 days ago is showing some growth! It's white, right below the injection port, and nothing smells weird.

Without Flash;

With Flash;

I'm scared that it's just more of that fucking cobweb bullshit. Time will tell. The other jars are not showing anything yet, but to be fair I was kinda absent minded when I inocced and got alot of the solution in the middle of the jars rather then up against the glass like the tek suggests, because I am stupid and forgot to hold the needle straighter.

Update 1/6/14 *
Went ahead and trimmed the fat of the OP since alot of it was no good and useless to the log. Focusing only on the equipment and the current BRF jars. Below is a pic of the best looking jar aka Jar A.



All of them are showing growth but this one is definitely taking the lead. Colonization rates are as follows;

Innoc on 12/22 Growth on 1/6 @: Jar A - 37%, Jar B - 15%, Jar C - 7%, Jar D - 2% Jar E - 0%(in the fridge ran out of spore juice in case you were wondering where #5 was)

However..., I did kinda botch the position when I was inoculating a few times which leads me to believe that alot of the colonizing on some of the other jars might be going on more at the core right now, for example, I only just barely started to see growth coming to the glass on D today.

I have another thread to track the daily growth of my jars mycella and advice as to whether it's developing safely or not. If you're interested you can find it here. More updates coming soon.

Update 1/18
Quote:

Camtaro420 said:
Quote:

Camtaro420 said:
Hey Hey everyone.

So as many of you know I have had hell the past month dealing with syringes. My last attempt was with a few jars of BRF done in classic PF tek style (only I used a PC instead of a steampot).

The fastest growing jar is starting to smell funky. Little bit like moldy I guess? It's not gag worthy but it definitely doesn't smell like the verm(that dirt like smell) used too and it doesn't smell like mushrooms either.

I Inoced on 12/22. Here is the pictures of the growth.

Updated 1/17 Jar A


Updated 1/18
Top view


Side 1 (where the hole that sells the most pungent of the 4 is)


Side 2 (Still waiting, a little closer every day =D)


Side 3


Side 4



Any words of advice? Is it normal for the verm barrier to smell a little weird towards the end? IS it just the GE? I'm a noob. No idea what's up.




Updated the OP. My Jar is starting to smell a little weird





Edited by Camtaro420 (01/18/14 08:27 PM)


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InvisiblePestile

Registered: 05/02/13
Posts: 875
Loc: Northern Europe
Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Camtaro420]
    #19277702 - 12/15/13 04:16 AM (10 years, 2 months ago)

Great project, will this be "from print to print" or are you using other methods of inoculation?
Beef agar sounds interesting, I will be waiting to hear how that works out for you. I've seen a beef agar recipe somewhere on this site, but never actually read any threads where it has been used.


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


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OfflineCamtaro420
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Pestile]
    #19277712 - 12/15/13 04:21 AM (10 years, 2 months ago)

Quote:

Pestilence said:
Great project, will this be "from print to print" or are you using other methods of inoculation?
Beef agar sounds interesting, I will be waiting to hear how that works out for you. I've seen a beef agar recipe somewhere on this site, but never actually read any threads where it has been used.




It will be close to print to print I will be doccumenting my work and I will continue to use spores, but I will be cloning and isolating too.I'm pretty excited to see the growth vs my own agar as well. 
:thisisgonnabegood:


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Edited by Camtaro420 (12/15/13 04:27 AM)


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OfflineUltron
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Pestile]
    #19277719 - 12/15/13 04:23 AM (10 years, 2 months ago)

:thisisgonnabegood:


--------------------
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OfflineCamtaro420
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Ultron]
    #19277784 - 12/15/13 05:01 AM (10 years, 2 months ago)

Well the pressure cooker has been cooking for 90 minutes at/around 15 psi. Turned off the burner. Falling asleep. In the morning I will take some pics. Glove up, wipe down, flame hot and put in about .5 CC in about  17 petris.

Since the syringe won't be touching anything after flaming should I bother and go;

A: flaming it again? (Just get all the petris ready to open, Flame the syringe once, then open 1, squirt close 1. Open 2 squirt, close 2, etc.)

or

B: Get petri 1 ready to open (remove the jar ring but not the top) -> Flame the syringe - > open and squirt in Petri 1 -> Close. Get petri 2 ready to open (remove the jar ring but not the top) -> Flame the syringe again - open and  squirt into syringe 2 -> rinse repeat and continue till all jars are complete with flame inbetween each one.

I want to say B but it seems like alot of air movement in and out of the SAB, and unlike BRF jars where you stick a syringe into dirt, I'm just squirting into open jars in a SAB environment.


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InvisiblePestile

Registered: 05/02/13
Posts: 875
Loc: Northern Europe
Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Camtaro420]
    #19277796 - 12/15/13 05:11 AM (10 years, 2 months ago)

I'd probably go with something in between, maybe flame every 3-4 dishes/jars and take it from there.

I hope you mean .5cc all in all. 1 drop from a spore syringe is quite enough to inoculate a agar plate. :smile:


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InvisiblebodhisattaMDiscordReddit
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Pestile]
    #19278253 - 12/15/13 08:53 AM (10 years, 2 months ago)

Quote:

Pestilence said:
I'd probably go with something in between, maybe flame every 3-4 dishes/jars and take it from there.

I hope you mean .5cc all in all. 1 drop from a spore syringe is quite enough to inoculate a agar plate. :smile:



:whathesaid:

just use a very small drop, and flame every few dishes, not every single one.

Also,
remove the vendor name in the original post, don't use a vendor name in conjunction with cultivation, it can get the vendor in trouble.

Lastly,
Using a hammer as weight on your PC scares the shit out of me, I would try to get one that works as intended asap.

Everything else looks good. Next time if you're making no pour agar jars though you can just put boiled agar right into the jars and PC them, You don't have to PC the agar first then PC the jars a second time. Just PC the jars and agar at the same time.

If you're pouring pre-sterilized petri dishes that you bought you PC the agar first and then pour it into the dishes in the SAB (like RR does in the agar video) then those plates are ready to go(unless you want to wait a 3-4 days to see if any contams show up before using them)


:thumbup:
:popcorn:


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OfflineCamtaro420
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: bodhisatta]
    #19279129 - 12/15/13 12:22 PM (10 years, 2 months ago)

Quote:

bodhisatta said:
Quote:

Pestilence said:
I'd probably go with something in between, maybe flame every 3-4 dishes/jars and take it from there.

I hope you mean .5cc all in all. 1 drop from a spore syringe is quite enough to inoculate a agar plate. :smile:



:whathesaid:

just use a very small drop, and flame every few dishes, not every single one.

Also,
remove the vendor name in the original post, don't use a vendor name in conjunction with cultivation, it can get the vendor in trouble.

Lastly,
Using a hammer as weight on your PC scares the shit out of me, I would try to get one that works as intended asap.

Everything else looks good. Next time if you're making no pour agar jars though you can just put boiled agar right into the jars and PC them, You don't have to PC the agar first then PC the jars a second time. Just PC the jars and agar at the same time.

If you're pouring pre-sterilized petri dishes that you bought you PC the agar first and then pour it into the dishes in the SAB (like RR does in the agar video) then those plates are ready to go(unless you want to wait a 3-4 days to see if any contams show up before using them)


:thumbup:
:popcorn:




Word. Excited to innoc after my wake and bake. The pressure cooker is still faintly warm though like as warm as a thin blanket would be after you took it off of yourself. Should I innoc using them that warm or should I refrigerate the dishes for awhile first.

As for the pressure cooker, lol, don't worry, it has all the pieces it's suppose to, just the pressure won't go past 10 psi on it's own with how old the weight valve is. Also it's a design, make and model I cannot find replacement parts for because it's so ancient.

My mother in law used to do the same thing and went to 18 back in her day, she just said to find something decent to weigh the excess pressure down with. I used a hammer. It climbed very slowly to 15 psi and then maintained well when the heat was slightly lowered. Just in case I had those knobs tight as all hell AND some vice grips on that lid.

And only 1 drop? I figured that was like .5 CC.


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InvisiblebodhisattaMDiscordReddit
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Camtaro420]
    #19279187 - 12/15/13 12:32 PM (10 years, 2 months ago)

You should find the AGAR prep videos part of the let's grow mushroom video series. That will answer your questions more ideal then I could type out. It will explain how to pour hot agar, how to make sure it's liquid before you pour, how now to burn yourself, how long to wait to inoculate agar plates, the benefits of letting them sit for a few days to make sure your plates are contam free, etc...

.5CC has a lot of drops


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OfflineCamtaro420
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: bodhisatta]
    #19279344 - 12/15/13 01:07 PM (10 years, 2 months ago)

If I seriously have that many drops to work with, what should I do with all the rest of syringe? I have like 13 jars with PDYA and 5 with BPA.

And while waiting would be safer for the contaims, since I PC it before and after and used SAB in-between I think I am safe. I just was wondering if you all think I should innoc them cold from the fridge or the sunny desert day room temp they are now.

The syringe is in the fridge. Should I let it warm to room temp first?


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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Camtaro420]
    #19279356 - 12/15/13 01:11 PM (10 years, 2 months ago)

Quote:

Camtaro420 said:
If I seriously have that many drops to work with, what should I do with all the rest of syringe? I have like 13 jars with PDYA and 5 with BPA.

And while waiting would be safer for the contaims, since I PC it before and after and used SAB in-between I think I am safe. I just was wondering if you all think I should innoc them cold from the fridge or the sunny desert day room temp they are now.

The syringe is in the fridge. Should I let it warm to room temp first?




make cakes and use the syringe on that or leave it in the fridge and make 100's of isolates from it over the years

You're probably right about being safe with the jars but if you're pouring dishes you might not have 100% success the first few times, it gets easier and eventually you'll probably like them better than working with jars.

You can inoc them cold or room temperature, no big deal it's going get back to room temperature pretty quick when you incubate them(room temp is fine for incubation)

You don't have to warm the syringe up.


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OfflineCamtaro420
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: bodhisatta]
    #19279376 - 12/15/13 01:16 PM (10 years, 2 months ago)

Word. I support that waiting on the poured agar. saving the syringe huh? that sounds risky but not to bad an idea

When flaming the syringe should I hold it with the syringe level, upwards, or downwards


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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Camtaro420]
    #19279393 - 12/15/13 01:22 PM (10 years, 2 months ago)

Quote:

Camtaro420 said:
Word. I support that waiting on the poured agar. saving the syringe huh? that sounds risky but not to bad an idea

When flaming the syringe should I hold it with the syringe level, upwards, or downwards




you can hold on to a syringe for years, I hold my syringe slightly upwards when I flame so I don't melt the luer-lock part of the needle.


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OfflineCamtaro420
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: bodhisatta]
    #19279408 - 12/15/13 01:28 PM (10 years, 2 months ago)

So when i'm done I'll just alchol wipe the plug, let it dry, unscrew the syringe head, then screw the plug back on?


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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Camtaro420]
    #19279414 - 12/15/13 01:32 PM (10 years, 2 months ago)

Quote:

Camtaro420 said:
So when i'm done I'll just alchol wipe the plug, let it dry, unscrew the syringe head, then screw the plug back on?




I leave the needle attached and put a needle cap over it, or put it in a baggie or saran wrap etc.. then into a small box and into the fridge.


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OfflineCamtaro420
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: bodhisatta]
    #19279437 - 12/15/13 01:37 PM (10 years, 2 months ago)

=o wont stuff come out of the needle!?


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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Camtaro420]
    #19279442 - 12/15/13 01:38 PM (10 years, 2 months ago)

Quote:

Camtaro420 said:
=o wont stuff come out of the needle!?




spontaneous plunger pressing? a drop or two no big deal


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OfflineCamtaro420
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: bodhisatta]
    #19279457 - 12/15/13 01:41 PM (10 years, 2 months ago)

Hrm you make a good point. So the cap. Do i need to clean that? I think it comes with a pre sterile one actually.

Innocing soon then..


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


Edited by Camtaro420 (12/15/13 01:41 PM)


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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: Camtaro420]
    #19279469 - 12/15/13 01:45 PM (10 years, 2 months ago)

Quote:

Camtaro420 said:
Hrm you make a good point. So the cap. Do i need to clean that? I think it comes with a pre sterile one actually.

Innocing soon then..




I wouldn't worry too much about how sterile the syringe going to be. You'll flame the needle next time any way and nothing is going to go through the plastic or down the needle. Besides, ideally you're going to want to put the spores directly to agar next time anyway.


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OfflineCamtaro420
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Re: Cam's grow #2. Agar, WBS, G2G, Monotub Tek [Re: bodhisatta]
    #19279478 - 12/15/13 01:47 PM (10 years, 2 months ago)

So like there's a huge clump of spores. Even tho I shake it and can't really get it to break up that's cool right cuz the millions of spores and all?


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