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Offlinecoult45
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First couple tries, not seeming to work out
    #23683105 - 09/27/16 12:36 PM (7 years, 5 months ago)

Hi folks, I'm new, this is my second try at growing. My first was last october-december, my BRF jars never seemed to colonize... anyways, heres what I did this time, and where I'm at


I have followed every single step as closely as possible with RR's videos and other PF teks.

I've used a pressure cooker, I've gotten my water/brf/verm mixture as accurate as possible. I've cooked it for 90 mins, and let it cool 12 hrs. I've used micropore tape and injected in a clean environment. I just can't see any sort of growth, and it has been a while.


Possible issues may include

Ambient temp is at the very lowest 60, the highest 70. I know this will slow things down, but they should still colonize right?

Also, what is the procedure of flame treating the needle, as well as wiping with alcohol? I would flame treat till hot, let it cool a few seconds, inject, and then wipe clean. This is correct, correct? The needles will sizzle when flame treated, is the also right? I have a feeling I'm killing the spores?

My spores are all from ***reputable dealer *** and I have 3 different types.

Here's the list of my timing of batches.

BRF Cakes - Half pint jars. 2:1:1 ratio Verm, water, brf, or perhaps just a hair less water


BATCH 1 - inoculated 6 jars ( b+) on August 29th 8pm, 1cc

BATCH 2 - inoculated 5 jars of Australia sept 1st 2 pm 1cc


Liquid culture creations: 3/4 tablespoon honey/maple mix in a half pint jar

#1 Inoculated 9/10 10am with 2 ml of B+
#2 Inoculated 9/10 10am with 2 ml of Cambodian



None of my jars, nor my LC's have any growth at all... bad syringes? bad procedures? too cold? wait even more?

Edited by coult45 (10/05/16 10:14 AM)

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Re: First couple tries, not seeming to work out [Re: coult45]
    #23683184 - 09/27/16 01:16 PM (7 years, 5 months ago)

remove where you got them from please.
read the rules.

also 2ml in a half pint is a touch much. you barely need half a ml.

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Offlinemrmazdarx9
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Re: First couple tries, not seeming to work out [Re: mushboy]
    #23683303 - 09/27/16 01:56 PM (7 years, 5 months ago)

You need 1/4cc per jar:thumbup:
One drop per hole.
I usually do 1/4cc per hole but have recently been told I was using more than needed.


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Offlinecoult45
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Re: First couple tries, not seeming to work out [Re: mrmazdarx9]
    #23686011 - 09/28/16 09:59 AM (7 years, 5 months ago)

Thanks for the input everyone. I changed my original post. In the liquid cultures, I thought I read more = faster germination, so I did 2 ml just to make it go quicker. Is this too much?


For the brf cakes, I did a 1/4 cc per hole, 4 holes in each lid, so each jar had 1cc. Is this too much? The above post says 1/4cc per jar, but I must be mistaken in doing 1/4 cc per hole....

Edited by coult45 (09/28/16 10:01 AM)

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InvisibleJacobStorm
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Re: First couple tries, not seeming to work out [Re: coult45]
    #23686069 - 09/28/16 10:17 AM (7 years, 5 months ago)

1/4 CC per jar will even work. I stretched a 10CC syringe over 20 jars. Dont stress adding too little. But anything over 1 CC a jar is just waste and counter productive.


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Offlineleech911
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Re: First couple tries, not seeming to work out [Re: mrmazdarx9]
    #23686123 - 09/28/16 10:38 AM (7 years, 5 months ago)

Quote:

mrmazdarx9 said:
You need 1/4cc per jar:thumbup:
One drop per hole.
I usually do 1/4cc per hole but have recently been told I was using more than needed.




So you've been told to apply 1/4 of 1/4 of 1 cc to every hole? Its hard enough to squeeze out just 1/4 of a CC per hole. I couldn't imagine trying to eyeball 1/16th of a CC per hole. 1/4 of a cc per hole i.e. 1 cc per jar should be fine. Even if you did do less you shouldn't be hurting the cake, it only takes 2 spores to make a colony!

when you flame sterilize you should hear sizzling this is completely normal, it's even normal for a little to shoot out when heating it up. I wouldn't be worried about killing the spores, as soon as you fill the needle with liquid it cools and anything coming out after has healthy spores. If it's been 30 days and 0 sign of growth I'd chuck it and start over.

Normally I do a 3:1 verm to BRF in PF jars but Ive done 1:1 and it still worked just took forever to colonize as the cake was mostly flour. on that note even 1:1 brf to verm was fully colonized in 30 days time.

Much luck with future endeavors, it may help us to help you if you post pics of your set up; but, based on what you've said, this is the best advice I can give you.


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InvisibleJacobStorm
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Re: First couple tries, not seeming to work out [Re: leech911]
    #23686131 - 09/28/16 10:44 AM (7 years, 5 months ago)

Quote:

leech911 said:
Quote:

mrmazdarx9 said:
You need 1/4cc per jar:thumbup:
One drop per hole.
I usually do 1/4cc per hole but have recently been told I was using more than needed.




So you've been told to apply 1/4 of 1/4 of 1 cc to every hole? Its hard enough to squeeze out just 1/4 of a CC per hole. I couldn't imagine trying to eyeball 1/16th of a CC per hole. 1/4 of a cc per hole i.e. 1 cc per jar should be fine. Even if you did do less you shouldn't be hurting the cake, it only takes 2 spores to make a colony!

when you flame sterilize you should hear sizzling this is completely normal, it's even normal for a little to shoot out when heating it up. I wouldn't be worried about killing the spores, as soon as you fill the needle with liquid it cools and anything coming out after has healthy spores. If it's been 30 days and 0 sign of growth I'd chuck it and start over.

Normally I do a 3:1 verm to BRF in PF jars but Ive done 1:1 and it still worked just took forever to colonize as the cake was mostly flour. on that note even 1:1 brf to verm was fully colonized in 30 days time.

Much luck with future endeavors, it may help us to help you if you post pics of your set up; but, based on what you've said, this is the best advice I can give you.




The proper and most prefered BRF ration is 1:1:2 water/BRF/verm.
I wouldnt fuck with this recipe its only going to be counter productive.

Anymore when Im inoculating BRF jars I just watch how much liquid I see running down the sides of the jar. Its how I measure a 1/8th of a CC


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Offlinecoult45
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Re: First couple tries, not seeming to work out [Re: JacobStorm]
    #23686216 - 09/28/16 11:12 AM (7 years, 5 months ago)

Weird. Last fall when I gave this a go, my mixture seemed a bit too wet, as in you could see "humidity" in the jars. Those colonized between 20-60%, but it also took a decent amount of time, probably 4 weeks to get to that stage. It's pretty darn dry where I live, indoors probably 15-30% all the time. Any other ideas?

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Offlinemrmazdarx9
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Re: First couple tries, not seeming to work out [Re: coult45]
    #23686269 - 09/28/16 11:31 AM (7 years, 5 months ago)

Theres always moisture in the jars when colonising


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InvisibleJacobStorm
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Re: First couple tries, not seeming to work out [Re: mrmazdarx9]
    #23686594 - 09/28/16 01:04 PM (7 years, 5 months ago)

Your RH in your house doesnt matter much when fruiting and certainly doesnt matter while colonizing.

If you followed the 1:1:2 ration and your stuff was too wet it was a failure on your part. That ratio is perfect in everyway.


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azur said: If you like 6th grade results, then 6th grade projects are great.

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Offlinecoult45
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Re: First couple tries, not seeming to work out [Re: JacobStorm]
    #23709456 - 10/05/16 10:13 AM (7 years, 4 months ago)

I think it is time to toss the brf jars. Still nothing.


On another note, LC #1 had a inoculation hole, using b+, like one of the sets of BRF and has no growth.

LC #2 was inoculated with cambodian, which I haven't put into any cakes at all, and has a much larger inoculation hole than all of my brf jars and is a bigger hole than LC #1 jar. My current phone has a really bad camera, so no pics today... but to me, and my research, it looks like some quality growth. It went slow, but its been 60 degrees in the room...


So....perhaps my holes in LC #1 and all my BRF jars are too small, not allowing any fresh air exchange. this is quite important isn't it? Perhaps this was my issue?

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Offlinemrmazdarx9
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Re: First couple tries, not seeming to work out [Re: coult45]
    #23709465 - 10/05/16 10:17 AM (7 years, 4 months ago)

By inoculated LC are you saying you used a spore syringe? If so that's your problem with the LC


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Offlinecoult45
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Re: First couple tries, not seeming to work out [Re: mrmazdarx9]
    #23709496 - 10/05/16 10:31 AM (7 years, 4 months ago)

Quote:

mrmazdarx9 said:
By inoculated LC are you saying you used a spore syringe? If so that's your problem with the LC





I understand that the spore syringe is not the best way to start a liquid culture. I started LC's in hopes that I could have some germinated myc ready to blast into BRF jars, as well as using the spore syringe straight into BRF jars. Couple different methods.


So either, my air exchange isn't happening in my BRF jars

or..

I think this is a far off thought, that perhaps the Cambodian strain is legit and the others are bunk...

I'm thinking my air holes are too small.





WAIT! I've left my micropore tape on after inoculation.... Should I have taken it off? Directly after innoc?

Edited by coult45 (10/05/16 10:33 AM)

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InvisibleJacobStorm
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Re: First couple tries, not seeming to work out [Re: coult45]
    #23709499 - 10/05/16 10:33 AM (7 years, 4 months ago)

Gas exchange isnt the problem. Try sticking to the tek you picked. Its all this improvising with stuff you no nothing about that has hindered your efforts.


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Ethnobotanical garden forum

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azur said: If you like 6th grade results, then 6th grade projects are great.

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Offlinecoult45
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Re: First couple tries, not seeming to work out [Re: JacobStorm]
    #23709529 - 10/05/16 10:44 AM (7 years, 4 months ago)

Quote:

JacobStorm said:
Gas exchange isnt the problem. Try sticking to the tek you picked. Its all this improvising with stuff you no nothing about that has hindered your efforts.




I have not improvised about anything. I followed RR's BRF tek. 2:1:1 ratio. Watching the videos, taping holes, everything.

On a completely separate note, I followed a different tek for liquid cultures.

I have yet to improvise a tek or method, I am simply asking questions about specific steps in the procedure.




Here's my point. Lets pretend I haven't tried any LC's at all...



At this point I've only done BRF cakes and they show no growth. 2 different syringes(b+ and Australian), 11 jars total. Followed the tek, 2:1:1, cooked them for the correct time and pressure. I was super careful about being sterile. No growth at all. Something isnt working, correct?

Now, as a side experiment, I have 2 LC's going. One has a hole larger than the other, and that larger hole has a syringe that hasn't been used yet. This one shows obvious growth (cambodian), the other, smaller and already attempted syringe (b+) shows nothing.


correct me if im wrong, but something is either wrong with my B+ and Australian syringes and not with my Cambodian

Or

The size of the Innoc port has something to do with it

Or

somehow my technique was different between the two, which I can tell you, I've measured everything as accurately as possible and kept the system consistent, so this shouldn't be the case.





It seems as though the micropore tape brings up all different views as far as using it vs not using it, removing after innoc and not removing. everyone seems to think their method works. My holes are tiny in my BRF jars, to the point at which the needles i bought for LCing (15 gauge), probably won't fit through the holes, but the spore syringes I bought from the sponsored seller do fit, because they are smaller...

Edited by coult45 (10/05/16 10:49 AM)

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Offlinemrmazdarx9
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Re: First couple tries, not seeming to work out [Re: coult45]
    #23709611 - 10/05/16 11:16 AM (7 years, 4 months ago)

IME micropore make no difference apart from maybe a little slower colonisation but that could have been due to other issue. I've used both with and without can't say there's any difference. The holes are ment to be like a drill bit size I stabbed through mine with a fork with just one point the rest bent down lol those lids have gone in the bin from rust I just use tin foil now with holes.


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Re: First couple tries, not seeming to work out [Re: mrmazdarx9]
    #23709637 - 10/05/16 11:27 AM (7 years, 4 months ago)

after re-reading the post your prob is LC

brf cakes do no like the extra water content of LCs IME atleast,
unlike grains which seem to soak it up. (again IME)

and its not that using a MS 'isnt the best way' its a terrible way.
even though spores are 100% contam free so they say, the are not free
of bacteria that will thrive in the LC.

pf tek is ment for MS spores. stick with that. :thumbup:

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