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Stickyicky247
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Registered: 09/09/15
Posts: 7
Last seen: 6 years, 29 days
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Some noobie help would be greatly appreciated
#22210614 - 09/09/15 02:41 AM (8 years, 4 months ago) |
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Hey guys! I finally lurked long enough to feel confident enough to try my hand at some mycology. I've researched for about a month now and feel I have at least a basic understanding. So the reason for my post is to see if I can be pointed in the right direction.
So here is what has been done:
Received a gt syringe and bought Verm, brf, half pint jars, quart jars, micro pore tape, a big (24qt?) pc, sterlite containers, silicone, cellulose filter discs, an outgrow Lc jar lid, 50lbs organic rye berries,organic honey, and a bunch of other odds and ends.
The plan: I am very impatient, and love a challenge. So I wanted to run some brf cakes while learning grain spawn. And as a backup fail safe, a jar of Lc to keep me supplied while I undoubtedly fail many projects while getting a handle on the learning curve of a noob attempting more advanced teks. I followed pf tek for brf cakes and prepped the jars making sure to follow to a tee. I punched 2 holes in the lids with a nail and then wrapped in tinfoil. After PCing at 15psi for an hour, I began making my glove box. I cut 2 holes and duct taped 2 long rubber gloves into sterilite container with lid being on the bottom. Now where I see a potential for failure is, I am not doing this at home so it's hard to be there when I need to be due to some schedule constraints. I returned 3 days later and resumed where I left off but all the while, I left jars in pc.I sprayed with 1:5 bleach solution, loaded jars, and set my syringe (which was refrigerated), lighter, and alcohol swabs, in there as well. Sprayed the works with bleach solution again and began after 10 mins. I followed procedure and sterilized needle, swabbed with alcohol wipe, and then removed tinfoil and performed my inoculation. Repeated same steps for rest of jars (12 total). I then loaded into my homemade incubator made of one large black tote with a smaller tote set inside. Underneath inner tub, I placed a seedling heating pad which got me to around 80-84 F. Placed jars and went to work on grain jars.... This is where I began banging my head against the wall... First time, I measured grain at 1 cup per quart jar and 200 ml water and then pc'd at 15 psi for 60 mins with filters in place. I also placed distiller water into quart jar with 500ml water to 2 teaspoons honey iirc, and placed Lc lid on. Wrapped all in tinfoil and then pc'd. Performed same sterilizing procedure as last time. I opened instead of directly injecting filter disk. I saved a little (.5ml) for Lc jar which I sterilized needle and alcohol swabbed injection port. Loaded that into chamber and waited.i ended up with some great growth in a few brf jars but ultimately ended up with purple contam before fully colonized. Ended up with 2 cakes ready to go. Rye jars all contaminated except for 1 and the grain smelled rotten or fermented is probably a better description. I think my moisture content was off as it appeared slightly wet. I tossed all but 1 which looked promising. I was a dummy and shook the jars at about 50% colonization hoping to cover more surface area and ended up with a lovely jar full of cobweb. Made terrarium for 2 cakes trying desperately to get some fruit. I cut 6, 2in holes and loaded breathable filter plugs into holes. Poured approximately 2 1/2 inches of perlite that was rinsed and then poured 3 cups of bleach water solution and set cakes on top of jar lids. That is currently awaiting results. I made another batch of rye. Tried desperately to maintain exact recipe and soaked 24hrs in fridge, then simmered while hungover and awoke an hr later. Loaded that into jars and noc'd up with some really great looking Lc. I followed sterile procedure but will elaborate if need be. Moisture content didn't seem bad. No pooling, not really wet to the touch but a few kernels would stick to my hand. I lifted lids again to inoculate. It's been approx 14 days with no sign of life but smells fermented again. So I made yet another batch of grain and this time it seems to look how I feel it should. But... It sat for 4 days in pc, again, due to availability issues. So is this substrate still viable? Also, no fanning is being performed due to lazy partner helping.. Well not helping. So I purchased an air stone and pump and hope to get by with that. I've got 5 syringes on the way of different strains. I'd like some help getting pointed into the right direction. I purchased a hepa tabletop purifier and garbage bags to try that as an easier glove box. I know there's a name for that but it escapes me. I also have Petri dishes en route to take on strain isolation as soon as I can figure out how to dial everything in. I'm currently at a loss as to where I'm going wrong. Planning on trying some more cakes just to attempt to get something while learning. Any and all help, criticism, and ideas are not only welcomed but encouraged! I can't wait to feel the success of seeing those beautiful pins start forming!! Thanks in advance!!
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Stickyicky247
Stranger
Registered: 09/09/15
Posts: 7
Last seen: 6 years, 29 days
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Re: Some noobie help would be greatly appreciated [Re: Stickyicky247]
#22210638 - 09/09/15 03:10 AM (8 years, 4 months ago) |
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Also, I have horrible add and see I left out many details. I ended up using micropore tape on brf cakes and only the ring and filter disk on grain jars. Idk if or where I'm messing up or if I'm just being impatient but I'm hoping someone can enlighten me. Thanks again ✌
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Re: Some noobie help would be greatly appreciated [Re: Stickyicky247]
#22210642 - 09/09/15 03:13 AM (8 years, 4 months ago) |
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I would like to help but... have you seen this wall of text? I can't get through it too far without my head and eyes starting to hurt.
Anything in particular you need to know? Not sure what resources you've been using but here's a dump.
http://www.shroomery.org/forums/showflat.php/Number/19140341#19140341 http://www.shroomery.org/forums/showflat.php/Number/20541405 http://www.shroomery.org/forums/showflat.php/Number/14217681 http://www.shroomery.org/forums/showflat.php/Number/19208976 http://www.shroomery.org/forums/showflat.php/Number/20048771#20048771 http://www.shroomery.org/forums/showflat.php/Number/13558505
If your pf jars were built to spec they won't need the micropore since the dry verm layer is the filter. Grain jars need a proper filter. I prefer synthetic filter discs over the cellulose ones, since they can get wet and survive many pc cycles.
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Stickyicky247
Stranger
Registered: 09/09/15
Posts: 7
Last seen: 6 years, 29 days
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Re: Some noobie help would be greatly appreciated [Re: Inocuole]
#22210651 - 09/09/15 03:21 AM (8 years, 4 months ago) |
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Lol I appreciate it. And yea, it's pretty excessive but I wanted to list everything I did so I didn't receive advice to do something I already performed. I guess mainly I'm wondering why I'm getting such poor growth and high contam rate. Thanks for the links btw.
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PabloSumgie
Stranger

Registered: 08/20/15
Posts: 84
Last seen: 8 years, 3 months
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Re: Some noobie help would be greatly appreciated [Re: Stickyicky247]
#22210710 - 09/09/15 04:25 AM (8 years, 4 months ago) |
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Ok I'm gonna try. Ask to explain further I'm gonna cover stiff breifly. Ok no nerd to wipe syringe after flaming. Incubator isn't necessary but that's debatable. I can't really help you with grains I use wbs. Uhm a glove box is kinda over kill look up a still air box much easier. Most will say look up a shotgun fruiting chamber but because of your availability I say look up the poor mans pod tek. I use it I place cakes and leave it for up to a week self contained positive pressure I like it. As for leaving grains in pc for long time you need to shake that shit up when its still kinda hot to redistribute the moisture then you can leave them. That could be your problem idk. One last thing in just like you and I wanted to try everything at once and it was overwhelming and wastes alot of money and I got small return. I'm saying keep doing what your doing but if all else fails go to the pf tek and a sgfc or pmp and master that first. The micropore tape isn't necessary but it certainly won't hurt besides maybe a little less gas exchange. And your grain lids drill a 1/4 inch hole in metal lid and silicone a price of the filter disk on top. The punch a smaller hole or two and silicone those over nice edge around hole for self healing injection port.
-------------------- Alright then, picture this if you will: 10 to 2 AM, X, Yogi DMT, and a box of Krispy Kremes, in my "need to know" post, just outside of Area 51. Contemplating the whole "chosen people" thing with just a flaming stealth banana split the sky like one would hope but never really expect to see in a place like this. Cutting right angle donuts on a dime and stopping right at my Birkenstocks, and me yelping... Holy fucking shit!
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xpsprogamer
TrippyTrauma

Registered: 07/17/15
Posts: 98
Loc: NorthEast U.S.
Last seen: 3 years, 3 months
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Re: Some noobie help would be greatly appreciated [Re: Stickyicky247]
#22210713 - 09/09/15 04:27 AM (8 years, 4 months ago) |
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for rye grains remember you have to soak it for 20 hours not in the fridge (you want to germinate some of the Bacteria so it rinses away easier. then bring to a boil in same water for the same reason for a like 10 mins and strain well until its dry to touch. a lot of dust and other particles coat the rye and if not soaked and rinsed properly contamination is more likely to happen. now this is just what ive experienced. I started with rye for my first time... ive gotten 4 grows under my belt now and have yet to have any contaminations. ive done probably 30 jars. and have fruited over an ounce. and have 3 trays about to knot for me. I have a first time grow log but its not super detailed as I am a noob. it all come down to sterile procedure. remember when removing the jars from the PC work in a small room like a bathroom. that has been Lysol'd and left to sit for 15 mins. and I recommend using the alcohol wipe before using the flame to sterilize. the alcohol wipe can contaminate the needle. these are just my opinions and by no means am I a professional. I have just seen some decent results. even with the mistake I made my first time. there is a link to it if u wana take a look. My First Grow
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NDStepp84
Stellar nuclear waste


Registered: 04/23/15
Posts: 4,956
Last seen: 1 month, 4 days
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Re: Some noobie help would be greatly appreciated [Re: xpsprogamer]
#22210855 - 09/09/15 06:17 AM (8 years, 4 months ago) |
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Also, never go spores>LC. You would want to go spores>agar do a few clean transfers then agar>LC, better yet agar>LI or agar>grains>grain to grain. Only one way to tell if LC is good, possibly waisting a jar and time testing it.
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"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do. -Robert A. Heinlein
Links and teks ND's grow log and discussion Plant thread
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Stickyicky247
Stranger
Registered: 09/09/15
Posts: 7
Last seen: 6 years, 29 days
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Re: Some noobie help would be greatly appreciated [Re: NDStepp84]
#22212411 - 09/09/15 02:07 PM (8 years, 4 months ago) |
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Awesome. Thanks for your help guys. I'm going to go see what everything is doing now. I'm definitely going to be trying a new approach. I have been wasting a lot of money on it but honestly, it's so interesting that I don't really mind the fails because I know once I get it down and perfected that having that skill is priceless!
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xpsprogamer
TrippyTrauma

Registered: 07/17/15
Posts: 98
Loc: NorthEast U.S.
Last seen: 3 years, 3 months
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Re: Some noobie help would be greatly appreciated [Re: Stickyicky247]
#22212530 - 09/09/15 02:34 PM (8 years, 4 months ago) |
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i wish you luck and that first fruit u see will make it all worth it in the end.
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spacechildo
proletarians rise up


Registered: 01/24/13
Posts: 19,243
Loc: Babylon
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Re: Some noobie help would be greatly appreciated [Re: Stickyicky247]
#22212548 - 09/09/15 02:37 PM (8 years, 4 months ago) |
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Quote:
Stickyicky247 said: and only the ring and filter disk on grain jars.
the grains will dry. make a 1/4" or so hole in the lid.
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xpsprogamer
TrippyTrauma

Registered: 07/17/15
Posts: 98
Loc: NorthEast U.S.
Last seen: 3 years, 3 months
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Re: Some noobie help would be greatly appreciated [Re: spacechildo]
#22212581 - 09/09/15 02:46 PM (8 years, 4 months ago) |
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Quote:
spacechildo said:
Quote:
Stickyicky247 said: and only the ring and filter disk on grain jars.
the grains will dry. make a 1/4" or so hole in the lid.
oh yes make sure u have both parts of the lid on and just get a 1/4 hole in it and fill it with the poly.
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NDStepp84
Stellar nuclear waste


Registered: 04/23/15
Posts: 4,956
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Re: Some noobie help would be greatly appreciated [Re: xpsprogamer]
#22213121 - 09/09/15 04:42 PM (8 years, 4 months ago) |
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Quote:
Stickyicky247 said: Awesome. Thanks for your help guys. I'm going to go see what everything is doing now. I'm definitely going to be trying a new approach. I have been wasting a lot of money on it but honestly, it's so interesting that I don't really mind the fails because I know once I get it down and perfected that having that skill is priceless!
With failure comes experience. Stick to the tried and true methods, keep that good attitude and you will go far. Here is a good write up on sterile technique: http://www.shroomery.org/forums/showflat.php/Number/21889950
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"I am free, no matter what rules surround me. If I find them tolerable, I tolerate them; if I find them too obnoxious, I break them. I am free because I know that I alone am morally responsible for everything I do. -Robert A. Heinlein
Links and teks ND's grow log and discussion Plant thread
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Stickyicky247
Stranger
Registered: 09/09/15
Posts: 7
Last seen: 6 years, 29 days
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Re: Some noobie help would be greatly appreciated [Re: NDStepp84]
#22381164 - 10/14/15 10:15 PM (8 years, 3 months ago) |
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Well, got some success to keep me motivated! Out of 30 some jars of some rye, and the rest BRF cakes, I ended up with 2 BRF cakes that weren't contaminated. Of those, 1 fruity nicely after i dunked and put a casing layer of verm around it. The other, grew what I suspect to be cobweb mold, or perhaps just a shitload of mycelium but didn't pin even after close to two weeks. So i discarded. Got 3 flushes off that little cake and then the caps started turning a grayish blue without any disturbance so I leaned more toward contam than bruising. Anywho, thanks for the help everyone! I know it's nothing spectacular but I feel like I accomplished something. Now currently, I have 10 jars of rye that I intend to use for spawn. I've done my homework but I'm curious in how to effectively isolate strains? I'm going to pour some dishes tonight to get started. Does anyone have advice on Using tetramycin antibiotic in the mix too help with bacterial contam? I bought a big bottle of it and it seems like it would work great. Just still a little nervous on the do's and don't(s) ya know? I don't want too create a biohazard haha
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RJ Tubs 202



Registered: 09/20/08
Posts: 6,016
Loc: USA
Last seen: 10 hours, 37 minutes
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Re: Some noobie help would be greatly appreciated [Re: NDStepp84]
#22381230 - 10/14/15 10:33 PM (8 years, 3 months ago) |
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Quote:
NDStepp84 said:
Also, never go spores>LC.
I'm curious why it's a poor strategy to go from spores to a LC
I've read that before. I think it' been said that spores are never totally sterile and that was the rationale... to use agar to get away from any contaminants.
BTW, can I ask another question without hijacking this thread? I'm sometimes lazy and use in-vitro pins from a PF cake (an unopened jar) to inoculate RGS or grain. Is there sometimes a chance that in-vitro pins on a cake are not totally sterile? Dont flame me for sometimes not cloning the traditional way, and using peroxide :-)
thanks
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Re: Some noobie help would be greatly appreciated [Re: RJ Tubs 202]
#22381446 - 10/14/15 11:44 PM (8 years, 3 months ago) |
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Quote:
RJ Tubs 202 said: Q: I'm curious why it's a poor strategy to go from spores to a LC A: spores are never totally sterile, use agar to get away from any contaminants.
Answered your own question. Why are you using peroxide? There's always a chance the fruit's not sterile, but chances are pretty good it is.
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newrook
Sucks at bulk



Registered: 03/20/15
Posts: 657
Last seen: 3 months, 5 days
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Re: Some noobie help would be greatly appreciated [Re: Stickyicky247]
#22381502 - 10/15/15 12:01 AM (8 years, 3 months ago) |
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Quote:
Stickyicky247 said: Well, got some success to keep me motivated! Out of 30 some jars of some rye, and the rest BRF cakes, I ended up with 2 BRF cakes that weren't contaminated. Of those, 1 fruity nicely after i dunked and put a casing layer of verm around it. The other, grew what I suspect to be cobweb mold, or perhaps just a shitload of mycelium but didn't pin even after close to two weeks. So i discarded. Got 3 flushes off that little cake and then the caps started turning a grayish blue without any disturbance so I leaned more toward contam than bruising. Anywho, thanks for the help everyone! I know it's nothing spectacular but I feel like I accomplished something. Now currently, I have 10 jars of rye that I intend to use for spawn. I've done my homework but I'm curious in how to effectively isolate strains? I'm going to pour some dishes tonight to get started. Does anyone have advice on Using tetramycin antibiotic in the mix too help with bacterial contam? I bought a big bottle of it and it seems like it would work great. Just still a little nervous on the do's and don't(s) ya know? I don't want too create a biohazard haha
Not familiar with the ant bac stuff but I'd do regular plates and work on bettering your sterile tek while using a SAB. If you have a colonized master grain jar you could open it quickly in a SAB and take out a single grain and drop it to agar.
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  THROW AWAY YOUR SGFC
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RJ Tubs 202



Registered: 09/20/08
Posts: 6,016
Loc: USA
Last seen: 10 hours, 37 minutes
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Re: Some noobie help would be greatly appreciated [Re: Inocuole]
#22381588 - 10/15/15 12:29 AM (8 years, 3 months ago) |
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Quote:
Inocuole said:
Quote:
RJ Tubs 202 said: Q: I'm curious why it's a poor strategy to go from spores to a LC A: spores are never totally sterile, use agar to get away from any contaminants.
Answered your own question. Why are you using peroxide? There's always a chance the fruit's not sterile, but chances are pretty good it is.
Thanks for your reply
The concept that syringe spores are not sterile is new to me. I just read it here the other day. Maybe I glossed over it during my younger pot smoking days!
I meant using peroxide in the more traditional way of selecting a nice fruiting body from a grow out and dipping it into peroxide, cutting it open, and removing a piece of tissue. Which I've done many times.
I usually use a LC to inoc grain or RGS, but recently things changed in my life and I don't have my PC or hood, so I did some direct syringe to grain inoc's (grain I got from a vendor here) this summer and nothing worked. I've wracked my head why. You think this is the reason? Grain stinks when it's contaminated and slimes out. It's much harder to tell when RGS is contaminated . . .
So . . . the reason why BRF cakes work is why? The mycelium outruns bacteria?
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Re: Some noobie help would be greatly appreciated [Re: RJ Tubs 202]
#22381614 - 10/15/15 12:36 AM (8 years, 3 months ago) |
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More or less. Most grows done for the longest time all had tons of bacteria and everybody seemed to think it was normal. It really is, the amount of spores is just so great compared to any contaminants that it gets a foothold first. In liquid media though, bacteria will multiply quickly and fill up the whole LC before long.
I'm still amazed sometimes at the fact that PF tek actually works, since I've put spores to agar and seen what that looks like, I don't know exactly how the hell a cake can survive it, I guess some of them don't.
That all said it's not impossible to have clean spores. It's just very difficult and unlikely.
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RJ Tubs 202



Registered: 09/20/08
Posts: 6,016
Loc: USA
Last seen: 10 hours, 37 minutes
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Re: Some noobie help would be greatly appreciated [Re: Stickyicky247]
#22381632 - 10/15/15 12:42 AM (8 years, 3 months ago) |
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Quote:
Stickyicky247 said:
I was a dummy and shook the jars at about 50% colonization hoping to cover more surface area and ended up with a lovely jar full of cobweb.
Can someone chime in about this phenomenon?
If we shake a jar that is colonizing well, and then growth stops and the grain is clearly contaminated, it is not the act of shaking that created the contamination, right? I was warned by a vendor here recently about the mistake of kneading a bag too soon and that puzzled me. Shaking at 50% seems quite all right to me, Stickyicky247. I tend to prefer 30%, which I think is the range RR recommends.
The vendor said From my experience, you shouldn't knead until at least 70% colonization. At this point, the mycelium is definitely established enough to recover from the knead. Doing it any sooner than 70% is risky in my opinion.
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Kalistis


Registered: 09/06/15
Posts: 2,265
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Re: Some noobie help would be greatly appreciated [Re: RJ Tubs 202]
#22381646 - 10/15/15 12:47 AM (8 years, 3 months ago) |
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So how do you know it is cobweb? Pics?
I shook my last batch at close to 90%... and they did just fine.
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