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OfflineLittleMario
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Registered: 10/23/07
Posts: 4
Last seen: 16 years, 2 months
Trials and Tribulations of a Newbee
    #7625598 - 11/12/07 12:48 PM (16 years, 2 months ago)

Alright, so this is my first post on the forum.

I look forward to getting advice here and hopefully the community is full of people with the textbook knowledge and practical experience to help me see this thing through. Though mycology is a new field for me, I have a background in organic chemistry through my university and through the old Hive.

I recieved my spores from TLG, about a month ago, I got the Creeper 8x syringes deal. So far i've used two of them with little success.

My first four jars were BRF/vermiculite quarts following the specifications of the PFTek. These were inoculated by multispore injection with 2.5 cc's each of solution.

Sterilization procedures have included pressure cooking, flame sterilizing the needle tip, lots of lysol, lysol wipes, and an immaculately clean bathroom. They were left to cool overnight and the spore solution was poured down the side of the jars.

They have been left to incubate for about three weeks now. The air temperature inside my incubator, and the temperature inside of the jars, has been a fairly consistent 86, I have observed temperatures from 83 to 87, depending on the outside temperature and how much water volume has evaporated out. I understand i need to back the temp down about three degrees, but I haven't been worried about maintaining precise control.


Now, after three weeks, three of the four BRF jars have shown zero growth. Nothing, nada. No mycelium, no contamination, simply nothing has happened. I do have one jar that began showing signs of growth in the second week, and after three weeks is about 20% colonized with no signs of contamination. The progress of this jar continues and is steady, but it occurs to me that it's been a bit slow. It also occurs to me that a 1/4 success rate, given identical preparations of each jar, is unnacceptable and will not suit my needs.

About a week ago I started four WBS jars. Prepared by a 16 hour soak, followed by PC'ing 90 minutes, prepared with tyvek filters and injected in a clean bathroom. After a week I have no mycelial growth in any of the jars, and two jars that have begun a green mold at the very top.

Discouraged, I have started two jars of 4% Karo in dH2O today, although I will not have stirring, one of which is already in the incubator.

Aside from the two WBS jars that contaminated, likely due to sloppy technique, my problem really seems to be that these spores just don't want to grow for the most part with these multispore injections, and the growth that is taking place seems inordinately slow. I thought that 2.5 cc's would be more than enough to make this happen in a timely fashion.

I am considering taking my one BRF jar that is 20% colonized and cannibalizing it for grain to grain into WBS, dropping some of it in liquid culture too. because starting from the spores doesn't seem to be working too well.

This is a newbee question, but what the hell am I overlooking? My results have been very discouraging.. is what I describe just atypically poor performance, or is it really this hard to get multispore injections to work successfully and consistently?


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OfflineOphanim
The Molecule'sSpirit
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Registered: 10/01/07
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Re: Trials and Tribulations of a Newbee [Re: LittleMario]
    #7625626 - 11/12/07 12:54 PM (16 years, 2 months ago)

I'm a noob on my first grow as well, but I have a thought on the BRF jars that show nothing - did you shake the syringe before you inocced? Maybe it's possible you just injected sterile water into those jars.

I'm on my first grow and seeing extremely inconsistent results. All of my jars show some growth, but after 9 days, most of them show very minimal growth and three of them look like they're about 70%.


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Offlinesoul_psychonaut
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Registered: 09/04/07
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Loc: up north, Manitoba
Last seen: 15 years, 9 months
Re: Trials and Tribulations of a Newbee [Re: Ophanim]
    #7625713 - 11/12/07 01:29 PM (16 years, 2 months ago)

I was having very similar issues up until I started making LC.. I totally recommend you try making some up and shooting up your jars with LC. It really stretches the amount you get out of a single syringe!

Best of luck!!


--------------------
Anyone down for cuttin' open a TonTon tonight?


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Offlineurayasan
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Registered: 04/10/06
Posts: 111
Last seen: 16 years, 27 days
Re: Trials and Tribulations of a Newbee [Re: LittleMario]
    #7625798 - 11/12/07 01:51 PM (16 years, 2 months ago)

All I can really suggest is to build a still air/glove box, this should bring your contamination rate down to a low percent or eliminate it. The small trouble it will be to build one is nothing compared to the waiting and stressing you're currently having to endure.

Additionally, I doubt that your plan for "BRF to grain" will be successful whatsoever, if your contaminate load is such that you are being breached by needle-sized holes into your jars, you can be assured that opening those all the way up to transfer chunks around will not work.

Sorry :frown:

P.S.: As far as slow growth goes, I thought that was a common occurrence with "creeper," also I wonder what TLG stands for? Who are they, are they a Shroomery sponsor?


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Offlinec0_hush
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Re: Trials and Tribulations of a Newbee [Re: urayasan]
    #7625941 - 11/12/07 02:17 PM (16 years, 2 months ago)

i would drop the incubator temp as 86F in jar temp is where you want to be max, and that's achieved with 84F ambient incubator temp. Others who know alot better even suggest mid to high 70's... I read somewhere on here that 2.5cc for each cake is way too much and will result in negative effect. Each cake should do ok with 1cc.


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Offlinedrwatson
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Re: Trials and Tribulations of a Newbee [Re: LittleMario]
    #7626052 - 11/12/07 02:38 PM (16 years, 2 months ago)

I think you're incubation temp is way too high.  86 will probably give an internal temp of 90+ which means you may have caused a "thermal death" due to overheating?  i'm not sure but I know that temps like that will at least leave you wide open to contaminates.

A glovebox is easy to make, easy to clean, and loads safer.  Make one and use it.

The PF Tek uses 1/2 pint jar and can take upto 28 days to colonize.
You've done Quarts which are = to 4 x 1/2 pints so they will take a lot longer to colonize.  BRF/Verm takes longer than WBS.  WBS is more common in quarts and is quicker to colonize. 

Try your hand at a liquid culture.  there are lots and lots of teks just pick one and roll with it.  I made some LC 4 days ago and it's ready to use right now! 

You should find RR's Tek for Grain Prep.  That dude knows his shit trust the experienced ones - hell everything I say is from someone elses experienced mouth I just read through 100's of threads before joining!

Good luck! Don't give up! Don't give in!  Find your way and stick to it!:sun: :grin: :sun:


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Offlineprimordialoddity
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Registered: 08/15/07
Posts: 182
Loc: PNW
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Re: Trials and Tribulations of a Newbee [Re: drwatson]
    #7626161 - 11/12/07 03:01 PM (16 years, 2 months ago)

Yeah it's just obvious that failure is a part of this process, I'm a newb...i did a 5% successful pf grow a couple years back...now I have books and videos and am planning on LC-Rye-Cased 50/50...

I haven't even got past the LC! lol.

here's my link about failure:
I suck.


NEVER GIVE UP!


--------------------
...to unite the powers of the conscious and unconscious, of intelligence and emotion.

NOTE: All posts and/or messages from this account are 100% fictional and should not be taken literally.


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OfflineLittleMario
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Registered: 10/23/07
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Last seen: 16 years, 2 months
Re: Trials and Tribulations of a Newbee [Re: primordialoddity]
    #7627132 - 11/12/07 06:16 PM (16 years, 2 months ago)

I've got one colonized jar that looks like it could succeed, two that are a week old with no growth but still might have a chance, three that have done nothing at all, and two that have a little contamination and no signs of growth. I have this fear that i'm shooting 1 for 8

Yeah, right now I have my nifty fischer digi-thermometer sensor sticking down into the verm of a failed BRF jar and it's reading 86.4. Tommorow I think i'll pull it out, adjust the water back up and turn the temp down a smidge.

And seeing my first light greenish mold on the top grains, my eyes are open, i've learned my lesson, i'm building a fully sealed glove box this week.

Just how temperature sensitive are we talking anyway? Will 2-3 degrees too much or 2-3 degrees too little cause a no-growth? Will it cause a significant stall or slow growth? So it's better to err on the low side?

It is difficult for me to obtain spores, last time it involved a LONG drive and an out of state mailbox. Anyone who is looking to share the love and help keep the strains alive, please PM me, because I will do my best to make them thrive and will be more than willing once i'm up to prints.

TLG is a spore company, not on the list of sponsors i think. I got the special they are running where they give you twice as many syringes if you get Creeper. I figure this isn't a plug because of my terrible results. I sure would love it if I could blame this on the syringes.. has anyone found that syringes from different companies have variable results?

Hopefully the liquid cultures will have more consistent results.


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Offlineyepyepyep
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Registered: 09/05/07
Posts: 147
Last seen: 26 days, 8 hours
Re: Trials and Tribulations of a Newbee [Re: LittleMario]
    #7627205 - 11/12/07 06:30 PM (16 years, 2 months ago)

The syringes are questionable if you're getting minor germination at some points. That being said, it is probably more likely that you've had clumping which if you don't shake to disperse you don't get spores on every injection point.

Either way, I'm sure that if you try LC you will get a solution that will colonize consistently.

Good luck.


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Offlinec0_hush
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Re: Trials and Tribulations of a Newbee [Re: LittleMario]
    #7627224 - 11/12/07 06:35 PM (16 years, 2 months ago)

a few degree fluctuation wouldn't be bad if you weren't already redlining... People that have done this many times will tell you anything over low 80's will slow things down drastically and that seems to be what you're experiencing. Who knows if you did permanent damage to your jars but you really should turn the temp down to 78-82 double quick.


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OfflineFunkatron9000
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Registered: 10/03/07
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Re: Trials and Tribulations of a Newbee [Re: LittleMario]
    #7627243 - 11/12/07 06:38 PM (16 years, 2 months ago)

Quote:

has anyone found that syringes from different companies have variable results?


YES!
There are some pretty shitty places out there in the world! I have gone through my fair share of bad syringes, used to get them from SB and
it was something like 50% 50%... they double your order "for free"
but out of two syringes only one was ever OK. the other one i swear was toilet water. 

Point is, you have to go through a reputable vendor or get a print and make your own syringes. I actually just had a home made EQ MS syringe
of mine show growth faster than my EQ LC, lol, and the multi spore injection is still in the lead... on rye and on the WBS!

Try and get you some new spores if you can.. I know it's a pain for you, but if you get some good spores it'll all be worth it. :thumbup:


--------------------
You cannot yet touch my daughters awesome boobage
:jiggly:
You must fight to the death in the Breastriary of Nippopolis.




I thought Gene Wilder was cool BEFORE he was dead.


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OfflineLittleMario
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Registered: 10/23/07
Posts: 4
Last seen: 16 years, 2 months
Re: Trials and Tribulations of a Newbee [Re: Funkatron9000]
    #7627362 - 11/12/07 07:03 PM (16 years, 2 months ago)

Thanks guys,

I'm gonna take the incubator temp back immediately as a precaution. This being my first run, I really have no idea how sensitive the conditions are. Anyway, I hope that eventually I can relax, because it drives me nuts to baby this stuff, watching and waiting for shit to grow is even worse than trying to watch a distillation all the way through.

Does anyone ever try to re-inoculate no-growth jars? I mean, they are definitely sterile, there has been no sign of contamination after three weeks in an incubator.


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OfflineMolasses
sobriety with aside of what thefuck am i doing?
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Registered: 11/03/07
Posts: 564
Loc: Marijuanaville, Vermont
Last seen: 11 years, 5 months
Re: Trials and Tribulations of a Newbee [Re: LittleMario]
    #7627649 - 11/12/07 07:50 PM (16 years, 2 months ago)

i actually just tried re-knocking up 2 jars that didn't do anything, just cuz i figured using some lc was better than throwing the jars away.

i'd tell you the results if i could see anything yet, its only been like 2 days


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