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Offlinejust_curious
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Advice needed.
    #23093481 - 04/07/16 02:09 PM (7 years, 10 months ago)

So I tried MS to agar from an untrusted vendor, and bacteria grew right where I put the spore water droplet.(mistake number one)

I then ordered a cube print, and cube syringe, and oyster print and oyster LC syringe from a trusted vendor in an attempt to finally get things started.

I went to inoculate agar from a print (2 agar dishes per species). I did it on Monday. Nothing appears to be happening on 3 out of 4 agar. The growth that I do see is on a cube agar dish and is tiny, but I'm beginning to think it might be contaminated. My agar is in the mail, but I don't think it's going to get here any time soon.

What I would like to do, is prepare some BRF/Verm 1/2 pints, get those to colonize, and then put them to my grain. The issue is, although my jars are wide mouth, they have shoulders. I read a thread on here about opening a colonized jar, scraping off small crumbles of the BRF/verm, and transferring over to my 1 quart grain jars. I would inoculate the half pints with the cube spore syringes and oyster culture syringe.

I've got plastic lids with just SFD's, but am assuming I have to take the original lids, punching out 4 holes in each, and using Tyvek in order to inoculate with the syringes in the BRF.

Here is my concern:
1. will the transfer work if the jar colonizes
2. If the oyster is already an LC, could I get away with going straight to grain, and doing the cube to the BRF jars.
3. Are there any cons with a BRF to Grain transfer.
4. Am I thinking too negatively about spore syringe and LC syringe to grain, and should I just go ahead and inoculate the grain?

I obviously need to practice using an inoculation loop. I made my own, maybe I used too thick of wire for the loop? I flame sterilized the loop, put it to agar to cool, scraped the spores onto the loop, and then circled the loop on the agar. Maybe I put too many spores with the loop? I just really want to get something happening.

Once I finally get the grain jars going, I will be using CVG for the bulk substrate for the cubes, and straw for the oysters. The cubes will be grown in doors all the way through. The oysters will be colonized inside, and fruited outdoors (possibly in doors also the, but original plan was outdoors)

Any advice would be great. I'm so determined to get things going. I really want to be successful at this. As soon as my agar gets here, I will prepare it using the grocery store tek inside of a pasty plate and will continue practicing agar. Thanks everybody.

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OfflineNewZooReview
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Re: Advice needed. [Re: just_curious]
    #23093517 - 04/07/16 02:23 PM (7 years, 10 months ago)

It is tempting to squirt that syringe at anything that MIGHT work. I know that it makes you itch with anxiety to think about not getting started. Hold back. Take a breath. You don't want to overbuy syringes. Save some $$ for better things (like more agar).

Here is what I would do to keep progressing without blowing the syringes.

1) pf-tek jars are pretty good to get things started and they are smaller they will succeed of fail faster. I would do a 3 jars each--1cc per jar. If they colonize you can scrap a bit into agar to start isolating, p2g as you said, fruit them in an SGFC, or shred them with a cheese grater into some bulk substrate to do a minimono (the pics in my sig are from that approach).

2) I would keep the up with the agar though. With your syringes, don't bother with the loop for now. Just put a single drop from the syringe right in the middle of your plate. Let it sit covered for a while before you move the plates so that the liquid sinks-in or evaporates a bit. That ensures that you have plenty of concentrated spores to germinate. You need a sacrificial squirt to cool the needle after you flame it, but you can get agar done for less than .25CCs.

3) Then put your 1/2 fullish syringes in the fridge to see how these projects work-out.


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Offlinejust_curious
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Re: Advice needed. [Re: NewZooReview]
    #23093570 - 04/07/16 02:32 PM (7 years, 10 months ago)

I already purchased the syringes and they are sitting in my fridge. I wont be able to birth the cakes sue to the type of jars I have (I bought them for grain) what I planned on doing was inoculating the BRF jars (around 5 jars for each strain), letting it fully colonize, and then flame sterilizing a fork and use that to break up the colonized jars, and then dumping those crumbled pieces into the grains. I was just unsure because the cube is a spore syringe, and the oyster is LC

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OfflineNewZooReview
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Re: Advice needed. [Re: just_curious]
    #23093612 - 04/07/16 02:41 PM (7 years, 10 months ago)

Quote:

just_curious said:
If the jars fully colonize and you use a still air box (SAB) for your transfer, I think you will be Ok to P2G. Do you have a PC to sterilize the grain jars? If not, I would go spend $10 for 1/4 pint jelly jars and do the pf-tek first. Otherwise, why risk it?





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Offlinejust_curious
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Re: Advice needed. [Re: NewZooReview]
    #23093633 - 04/07/16 02:48 PM (7 years, 10 months ago)

Yes, I have a pressure cooker and SAB and alcohol lamp. So would both LC and MS syringes work with the PF tek

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OfflineNewZooReview
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Re: Advice needed. [Re: just_curious]
    #23093688 - 04/07/16 03:02 PM (7 years, 10 months ago)

Quote:

just_curious said:
Yes, I have a pressure cooker and SAB and alcohol lamp. So would both LC and MS syringes work with the PF tek



I don't know oysters but I had some left-over BRF/Verm mix and a bunch of kickass LC from a Puerto Rican variety so I squirted them together to see what would happen. I have a half-dozen jars that grew like crazy in 2 days. These aren't pf-tek. This is a pic after about 12 days. The BRF is just in a lump in the jar and the myc is thicker than I have ever seen it. Not sure what I will do with these things. Might try to dry them out for a long-term myc storage.



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InvisibleInocuole
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Re: Advice needed. [Re: NewZooReview]
    #23093889 - 04/07/16 03:53 PM (7 years, 10 months ago)

Do the ol... Use the red hot needle to make an indention in the agar to drop the spores trick.  Seems to help some people.


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                            :rainbowdrink: Tea doesn't work?                            AMU  (Q & A)                  Grain prep for Intergalactic Space Oats :pes:     

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Offlinejust_curious
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Re: Advice needed. [Re: Inocuole]
    #23094079 - 04/07/16 04:44 PM (7 years, 10 months ago)

Great, thanks. Do any of you put food coloring in your agar when you prepare it. I see pastywhyte uses it for his pasty plates and I want to, too. I only have the gel food coloring laying around the house. I was wondering if it would be possible to add a TINY bit of it to my agar. The only think that makes me second guess is the fact that it has corn syrup, sugar, and a few other things. I didn't want to add anymore sugar because 1: his recipe says to make sure your agar doesn't have any added sugars, 2: I will already be adding sugar via honey

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InvisibleInocuole
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Re: Advice needed. [Re: just_curious]
    #23094083 - 04/07/16 04:45 PM (7 years, 10 months ago)

Yeah you can add a single drop or so.  My food coloring is like a goop and has sugar in it, never bothers the mycelium any as far as I can tell.  I prefer blue but red is pretty popular too, and purple's a good halfway.  Green, maybe not so much, for obvious reasons.


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                            :rainbowdrink: Tea doesn't work?                            AMU  (Q & A)                  Grain prep for Intergalactic Space Oats :pes:     

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Offlinejust_curious
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Re: Advice needed. [Re: Inocuole]
    #23094094 - 04/07/16 04:48 PM (7 years, 10 months ago)


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Re: Advice needed. [Re: just_curious]
    #23094099 - 04/07/16 04:49 PM (7 years, 10 months ago)

Oh, god, no.  :lol:  I don't know about that.  Mine is in little dropper bottles from like a bakery shop, I think it's for cake icing.  I steal it from my lady.


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                            :rainbowdrink: Tea doesn't work?                            AMU  (Q & A)                  Grain prep for Intergalactic Space Oats :pes:     

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Offlinejust_curious
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Re: Advice needed. [Re: Inocuole]
    #23094111 - 04/07/16 04:52 PM (7 years, 10 months ago)

well it's not icing. It is just food coloring, no intended for eating plain. But I tried it and it has a hint of sugar. I just wanted to make sure it was no different then the liquid. It's just in gel form for decorating so it doesn't dilute into the product your putting it on...if that makes sense

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OfflineNewZooReview
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Re: Advice needed. [Re: just_curious]
    #23094112 - 04/07/16 04:52 PM (7 years, 10 months ago)

Should be fine. I starting using different colors to mark which recipe I used for the plate. I am trying different ones and you should rotate anyway. They build up in the fridge because I usually make more than I use so having an easy way to tell what they are is handy.


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Offlinejust_curious
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Re: Advice needed. [Re: NewZooReview]
    #23094149 - 04/07/16 05:01 PM (7 years, 10 months ago)

So you don't think the corn syrup, sugar, food starch-modified, citric acid, and sodium benzoate (preservative) would be an issue? I'm not sure the ingredients of liquid food coloring, but I'm wondering how similar it is. Maybe I will add a small dab to a tiny portion of liquid to get it into liquid form, and then add it to the agar. I'm hoping maybe Pasty will stop by :smile:

I've decided that I am not going to do the P2G. I will keep working agar until I have an isolate to work with and the go to grain. The reason I want to start the oysters is because it is only getting hotter and I want to grow outside. I bet I end up having to put my laundry basket into something that would hold ice, and then set the laundry basket on top of the ice. Just enough to bring down the temperature of the substrate and hopefully trick it to think its in its normal fruiting conditions...or maybe that sounds completely ridiculous, I have no idea lol

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Re: Advice needed. [Re: just_curious]
    #23094151 - 04/07/16 05:02 PM (7 years, 10 months ago)

A single little dab of anything won't fuck up your shit, just try it.  Even if it was pure sugar it would still work.  That's like, a drop per batch, not per plate, to be clear.  I'm almost positive mine has all that shit in it too.


--------------------
                            :rainbowdrink: Tea doesn't work?                            AMU  (Q & A)                  Grain prep for Intergalactic Space Oats :pes:     

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Offlinejust_curious
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Re: Advice needed. [Re: Inocuole]
    #23094184 - 04/07/16 05:07 PM (7 years, 10 months ago)

fuck it, I'll give it a shot. The gel is notorious for having a lot more powerful dye capabilities than liquid coloring

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Re: Advice needed. [Re: just_curious]
    #23094193 - 04/07/16 05:09 PM (7 years, 10 months ago)

Damn right it does.  Like I said, one drop does the trick for 8 plates for me, I do about a drop and a half for 16.  If it's even remotely the same you'll have good results.


--------------------
                            :rainbowdrink: Tea doesn't work?                            AMU  (Q & A)                  Grain prep for Intergalactic Space Oats :pes:     

Coir-ca-cola
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Offlinejust_curious
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Re: Advice needed. *DELETED* [Re: Inocuole]
    #23094235 - 04/07/16 05:18 PM (7 years, 10 months ago)

Post deleted by just_curious

Reason for deletion: figured it out


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InvisibleInocuole
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Re: Advice needed. [Re: just_curious]
    #23094421 - 04/07/16 06:00 PM (7 years, 10 months ago)

Turn up the heat..  Add a touch more water, just a few ml.  That happens to me sometimes.  Add the food coloring last, btw.  Probably too late..


--------------------
                            :rainbowdrink: Tea doesn't work?                            AMU  (Q & A)                  Grain prep for Intergalactic Space Oats :pes:     

Coir-ca-cola
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Offlinejust_curious
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Re: Advice needed. [Re: Inocuole]
    #23094603 - 04/07/16 06:43 PM (7 years, 10 months ago)

I added the food coloring and honey last. I did however forget to turn the heat down when my PC jiggled. It only went about 3 minutes though. bumped it down to low on my stove. Hoping they turn out okay.

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