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OfflineDurban.Poison
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Brf hasn't germinated
    #15858643 - 02/24/12 05:51 PM (12 years, 10 months ago)

I am a noob, with a couple of successful brf grows under my belt. I have previously bought pre-sterilized substrate jars, but for my last grow, I decided it was time to make my own jars. I used Bob's Red Mill brf, Sta-Green Horticulture Vermiculite. I followed RR Tek and inoculated with Koh Samui ms that I ordered from a sponsored vendor. It has been 12 days and no signs of germination/colonization.
1) One issue I did have is that I sterilized for 2hr15min, due to the fact that I fell asleep. Could this have been where I went wrong.
2) The syringe I received was totally clear, unlike all the others I have received in the past, which either had clumps or charcoal in them.

Any feedback will be greatly appreciated

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InvisibleKing of Pain
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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15858682 - 02/24/12 06:01 PM (12 years, 10 months ago)

whats the temps like where there stored? is there anything covering your innoc holes?

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OfflineDurban.Poison
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Re: Brf hasn't germinated [Re: King of Pain]
    #15858709 - 02/24/12 06:07 PM (12 years, 10 months ago)

I have them in a double tub incubator that holds between 74 and 80F...there are four inoculation holes that are not covered

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InvisibleKing of Pain
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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15858795 - 02/24/12 06:29 PM (12 years, 10 months ago)

well damn sounds like you know what your doing. Im stumped, could be a bunk syringe. Maybe that strain takes a long time to start.

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OfflineAmanita virosa
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Re: Brf hasn't germinated [Re: King of Pain]
    #15858935 - 02/24/12 07:20 PM (12 years, 10 months ago)

How deep is your verm layer?  are you sure you got the syringe below the verm?  the spores will not grow in the verm layer.  It is also possible  that over sterlizing dried out your substrate too much making it inhospitable to the spores.  Did you let the substrate cool properly before you woke up and shot the spores?  usually, if a syringe is bad, its because it is contaminated, not because its void of spores,but it is possible that you got a syringe with nonviable spores or lacking spores.

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OfflineDurban.Poison
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Re: Brf hasn't germinated [Re: Amanita virosa]
    #15859450 - 02/24/12 09:16 PM (12 years, 10 months ago)

the verm layer was 1/2 in. to 3/8 in. and I am sure I inoculated below this layer. I let it cool for about 27 hours before i used...I left them in the pot without removing lid until I was ready to inoculate...I'm guessing maybe I did dry them out, or maybe the verm I am using is very fine and I should have used more water in the mix. Should the brf mix be to field capacity?

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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15859470 - 02/24/12 09:20 PM (12 years, 10 months ago)

Being new to it as well i am in the same boat. no growth after 7 days... so i started over on the whole process tonight. gotta keep trying.  best of luck on your grow, mate. hope you get some signs of life.

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OfflineAmanita virosa
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Re: Brf hasn't germinated [Re: Shaddox]
    #15859601 - 02/24/12 09:41 PM (12 years, 10 months ago)

fine verm is good, but i suspect you cooked the water out of the substrate.  shaddox, give it two weeks before giving up on those jars

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Re: Brf hasn't germinated [Re: Amanita virosa]
    #15859631 - 02/24/12 09:46 PM (12 years, 10 months ago)

Quote:

Amanita virosa said:
fine verm is good, but i suspect you cooked the water out of the substrate.  shaddox, give it two weeks before giving up on those jars



:whathesaid:

PCing so long probably screwed your substrate moisture


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Re: Brf hasn't germinated [Re: Amanita virosa]
    #15859718 - 02/24/12 10:05 PM (12 years, 10 months ago)

Quote:

Amanita virosa said:
shaddox, give it two weeks before giving up on those jars




Oh, I have not given up on them yet. but i did order some more spore syringes from a sponsored vendor and went ahead and prep'd some new BRK cakes for when they arrive. Should be in tomorrow.


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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15859804 - 02/24/12 10:24 PM (12 years, 10 months ago)

I would say that you most likely dried out the substrate if it was in the P.C. that long.:confused:


--------------------
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We are a way for the Cosmos to know itself. -Carl Sagan

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OfflineDurban.Poison
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Re: Brf hasn't germinated [Re: silentshawn]
    #15860891 - 02/25/12 07:50 AM (12 years, 10 months ago)

I actually steam sterilized...not pressure cooked...does that make a difference?

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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15860955 - 02/25/12 08:27 AM (12 years, 10 months ago)

it shouldn't i steam mine.

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OfflineDurban.Poison
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Re: Brf hasn't germinated [Re: King of Pain]
    #15860974 - 02/25/12 08:33 AM (12 years, 10 months ago)

I have seen people before say that they steam sterilize for 2hr, so I didnt think it was that big of a deal that I went over 15min, unless the moisture content wasn't high enough when I put them in the jars, due to the very fine verm...It make sense to me that since the verm has more surface area that it would require more water, but I may be totally off on this...I really need to know if I should add more water next time, or if there is a way to tell if you got it right?

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OfflineAmanita virosa
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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15861006 - 02/25/12 08:46 AM (12 years, 10 months ago)

if you steam  sterilized it and not pcd it, AND you followed the basic pf tek recipe exactly,  you might have gotten a bum syringe.  Have you opened one  of the jars?  Inspected the  injection sites.  Might as well have alook at this point.  this is why if makes sense to test syringes on agar if  you have the means.  All you need is a SAB some agar and some plates.  That way you can "prove" whether or not a culture is what it is or not.

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Re: Brf hasn't germinated [Re: Amanita virosa]
    #15861031 - 02/25/12 08:54 AM (12 years, 10 months ago)

I really think I did get a bunk syringe, I just want to rule out the possibility that I made a mistake...My mom has an old microscope I was gonna try and get off her for this reason...So I can give it a look before I waste my time...I just need to come up with a reason that I need it...haha

Does anyone have any input on the moisture level of the mix...like should it be field capacity or should I just stick with the recipe

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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15862801 - 02/25/12 05:56 PM (12 years, 10 months ago)

I would stick with the recipe.  It is close to field capacity, but it depends on the %moisture of the BRF and verm when you started.  theres a little wiggle room in there so if you followed the recipe you must have had a bum syringe.

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OfflineDurban.Poison
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Re: Brf hasn't germinated [Re: Amanita virosa]
    #15863307 - 02/25/12 07:52 PM (12 years, 10 months ago)

Thanks for all yours and others help...it is greatly appreciated....I love this site

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Offlinekdmmontana
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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15865360 - 02/26/12 08:51 AM (12 years, 10 months ago)

Quote:

Durban.Poison said:
I have them in a double tub incubator that holds between 74 and 80F...there are four inoculation holes that are not covered




You dont need an incubator so you can take them out right now.

Quote:

Amanita virosa said:
fine verm is good, but i suspect you cooked the water out of the substrate.  shaddox, give it two weeks before giving up on those jars




2 weeks is fr to long for germination to take place. I have visible mycelium within 5 days and never had to wait any longer. If his jars dont germinate, I believe it to be because of the substrate blend/water content.

Quote:

Durban.Poison said:
I actually steam sterilized...not pressure cooked...does that make a difference?




Steam sterilizing is fine. You dont need to over do it. I cook my BRF substrates for about 1 hour and thats just fine. Boiling them for too long can cause them to become dry.  Also take care to cover your jars with foil properly.

Post up some pictures of your jars, I can bet my pension on that its the substrate.

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OfflineDurban.Poison
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Re: Brf hasn't germinated [Re: kdmmontana]
    #15865537 - 02/26/12 10:01 AM (12 years, 10 months ago)

I'll get back with those pics...asap...my camera is broke so I will have to borrow one from my buddy...but looking at them, there is an obvious color difference in the mix versus the dry verm layer and what looks to me to be an obvious moisture level difference as well, but I would love to see what you think

As for the incubator...my house stays at bout 40F to 60F this time of year, unless your 2 feet from the Dearborn...I  live in a old drafty house with no insulation...pretty sure the incubator is a must have

thank a bunch

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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15865546 - 02/26/12 10:04 AM (12 years, 10 months ago)

Sloshy substrate is caused by too much water and to fast work. Let the water sink into the verm for 10 minutes or so and then add your fine ground BRF bit by bit. Mix well.

Here is a picture of what youre looking for:





/KD

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OfflineDurban.Poison
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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15875054 - 02/28/12 07:50 AM (12 years, 10 months ago)

A quick update...I never got that camera, but I dumped the jars this morning and shed a little tear for them....to some of your surprise, not really mine, the moisture level in the jars was the same as when I put it in there....when i dumped the jars, the dry verm layer just poured off and the mix stuck in the jar...with a light tap they popped right out, but kept the form of the jar, until you broke it up by hand and it clumped nicely in my hand...you couldn't squeeze any water out, but you couldn't before I put them in the jar either...just clumps nicely...so what you think about that?

I would love your feedback before I proceed with my next set...I think it had to be a bunk syringe

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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15875195 - 02/28/12 08:48 AM (12 years, 10 months ago)

The substrate should not have alot of clumping. This is a indicator that your substrate was overly wet and probably the primary reason why germination was taking so long.

I like to hydrate the vermiculite to field capacity, tilt the bowl until I see a little bit of water pool and then add in the cup of brf for 3.5cups of verm.

Works perfect every time


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Re: Brf hasn't germinated [Re: HybridprX]
    #15875863 - 02/28/12 12:08 PM (12 years, 10 months ago)

wow..that's not like any recipe I heard before, but I bet it colonizes fast that way...does it flush out like normal?

anyways I started over and I followed the Tek recipe, as well as all the previous tips give by readers and hope I get this time

As far as bringing it to field capacity...I did a little test and I went 1 to 1 verm to water and it still wasn't at field capacity or any pooling in the bowl...not sure I am doing different, or maybe due to fine verm...and when I dumped the other jars there wasn't no clumping, just held the shape of the jar till you touched it...then it broke up easily

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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15876262 - 02/28/12 01:47 PM (12 years, 10 months ago)

you can test the water content by squeezing the verm. No more than a drop should come out. I bet HybridPrX knew that he just forgot to mention it. I always squeeze my mix after I added the BRF. If it stays a solid shape, its good, if it falls from my hand its too dry.

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Re: Brf hasn't germinated [Re: kdmmontana]
    #15876525 - 02/28/12 03:06 PM (12 years, 10 months ago)

thanks Montana...that's what I ended up with...if this dont work, I am getting some new verm

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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15876665 - 02/28/12 03:39 PM (12 years, 10 months ago)

I wanted to give a shout out to Lil' Shop of Spores for replacing the syringe in question, even though there is still some user error doubt, that I went over with them in detail...they have very great customer service...thank you very much!

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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15879825 - 02/29/12 05:24 AM (12 years, 10 months ago)

I should have added that when you hydrate the vermiculite mix it vigorously to ensure that it has absorbed as much water as possible. If you dont see roughly a spoonful and abit of pooling water then you need to add a few drops until you do. Then when you add the brf it will absorb the excess moisture and bring the substrate to perfect field capacity.


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OfflineDurban.Poison
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Re: Brf hasn't germinated [Re: HybridprX]
    #15879873 - 02/29/12 05:56 AM (12 years, 10 months ago)

thanks...I'll try that next time

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Re: Brf hasn't germinated [Re: Amanita virosa]
    #15879897 - 02/29/12 06:13 AM (12 years, 10 months ago)

Quote:

Amanita virosa said:
How deep is your verm layer?  are you sure you got the syringe below the verm?  the spores will not grow in the verm layer.  It is also possible  that over sterlizing dried out your substrate too much making it inhospitable to the spores.  Did you let the substrate cool properly before you woke up and shot the spores?  usually, if a syringe is bad, its because it is contaminated, not because its void of spores,but it is possible that you got a syringe with nonviable spores or lacking spores.




I don't think it would be possible to dry out your substrate from cooking too long, as long as you don't boil your PC/Pot dry.  As long as you have steam, you have a 100% humid environment outside your jars.  There's no where for that moisture inside the jar to evaporate off to - it would be like staying in a sauna so long you dried off.


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OfflineAmanita virosa
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Re: Brf hasn't germinated [Re: Stevaz]
    #15880048 - 02/29/12 07:30 AM (12 years, 10 months ago)

Quote:

Stevaz said:
Quote:

Amanita virosa said:
How deep is your verm layer?  are you sure you got the syringe below the verm?  the spores will not grow in the verm layer.  It is also possible  that over sterlizing dried out your substrate too much making it inhospitable to the spores.  Did you let the substrate cool properly before you woke up and shot the spores?  usually, if a syringe is bad, its because it is contaminated, not because its void of spores,but it is possible that you got a syringe with nonviable spores or lacking spores.




I don't think it would be possible to dry out your substrate from cooking too long, as long as you don't boil your PC/Pot dry.  As long as you have steam, you have a 100% humid environment outside your jars.  There's no where for that moisture inside the jar to evaporate off to - it would be like staying in a sauna so long you dried off.




Might be difficult with BRF/VERM but you can def cook the moisture out of grain if you over sterilize it.  I have done it before, esp with rye.  the kernels shrivel and dry.  Not sure about the physics of it though.

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Re: Brf hasn't germinated [Re: Amanita virosa]
    #15882030 - 02/29/12 04:41 PM (12 years, 10 months ago)

This brings up another question for me...since the spores are hydrated in water, and if you inoculate against the side of the glass, shouldn't you see some sort of colonization, regardless of the substrate moisture level...considering there were no contams and the spores were viable

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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15883573 - 02/29/12 10:26 PM (12 years, 10 months ago)

yes you will but that water wont last for long

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Re: Brf hasn't germinated [Re: kdmmontana]
    #15900914 - 03/04/12 08:38 PM (12 years, 10 months ago)

well I ditched the old jars and started over with a few experiments...the first batch of 5, I did the same way as before...the second batch of 5, I added more water till there was a little pooling, then went from there...the third batch of 4, I found some new, more coarse verm and was it much easier to deal with

I sterilized all three for 60 minutes and used the same syringe, so I think I have a pretty good test going...we will see which works better for me

I went a little lite on the ms on the first 2 batches, due to lack of confidence...about 3cc for 5 jars...the third batch, I went with a full cc per jar

I also tried my first lc...wish me luck

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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15902678 - 03/05/12 08:22 AM (12 years, 10 months ago)

If your using a pot to steam the pf jars 60 minutes is not sufficient, they require 90.

45-60 minutes if you're using a pressure cooker thats operating between 12-15psi.


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Re: Brf hasn't germinated [Re: HybridprX]
    #15903017 - 03/05/12 10:08 AM (12 years, 10 months ago)

Quote:

HybridprX said:
If your using a pot to steam the pf jars 60 minutes is not sufficient, they require 90.

45-60 minutes if you're using a pressure cooker thats operating between 12-15psi.




Ive steamed jars for 30 minutes and they were just fine. However those were 125 ml. The regular ones I usually steamed for an hour and that was fine. For a 500 ml substrate I would sterilize for at least 90 min. Thats just me, but you are correct:)

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Re: Brf hasn't germinated [Re: kdmmontana]
    #15928872 - 03/10/12 10:08 AM (12 years, 10 months ago)

just wanted to give an update...I check on my jars today and every inoculation point on every jar (except one had signs of growth)...55 out of 56...pretty good turn out...seems like the 3rd batch, with the medium grade verm, is progressing a little faster than the first 2 batches with the vine verm...it was done last and coming in first so far...I expected as much, but glad that I tested it out...I'll let you know if they actually reach 100% first or not...pretty sure they will, since I went a little heavier on the ms in that batch

what have I learned so far...moisture level doesn't matter much for seeing signs of growth when using a ms...achieving full colonization may be a different story though...time will tell

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Re: Brf hasn't germinated [Re: Durban.Poison]
    #15928923 - 03/10/12 10:26 AM (12 years, 10 months ago)

Quote:

"moisture level doesn't matter much for seeing signs of growth when using a ms...achieving full colonization may be a different story though...time will tell"




Excellent observation. Too much moisture will cause issues later on, I have myself experienced this and its true. Too much water is never good.

Give us pictures:)

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Re: Brf hasn't germinated [Re: kdmmontana]
    #15928983 - 03/10/12 11:01 AM (12 years, 10 months ago)

I figure I have a pretty good test going to see how much is too much, or not enough, or just what will work best for me....I definitely see running into some problems with at least one set, but that's what makes it fun...if everything went perfect all the time, that would be kinda boring to me...think I might add some gypsum next time to see if that changes thing much

A camera is definitely high on my list of must haves

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