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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7599696 - 11/05/07 06:46 PM (16 years, 2 months ago)

Here's a little myco porn for you guys. These are the left over mycelium water syringes from yesterday. Some people don't like sloppy seconds, but we're pretty clean so we figured we'd risk it.







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OfflineNationofStrangers
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7599758 - 11/05/07 07:00 PM (16 years, 2 months ago)

Go big or go home i guess~! nice work man.


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: NationofStrangers]
    #7599795 - 11/05/07 07:05 PM (16 years, 2 months ago)

heck yeah man, go big or go home, for no other reason than to do it. i've got one of those all or nothing personalities :bongload:


Edited by psilopod (11/05/07 07:10 PM)


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7600922 - 11/05/07 11:11 PM (16 years, 2 months ago)

Checked in on things and figured I'd post up a few pictures.  We did the grain to grain transfers only yesterday and they are already blowing up.  Here's some pics of the g2g about 24hrs after transfer. I wish I knew which strain is which off hand but I'm not sure.






  :bongload:


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7604929 - 11/06/07 09:51 PM (16 years, 2 months ago)

Well, it's day 2 since the first round of inoculation and things are going well. The g2g is progressing along at a satisfying pace and the spawn bags are showing their first signs of colonization which I don't think is too bad considering it's only been about 48 hours since inoculation.

Today was a busy day and we got quite a few things done, the biggest thing being the agar transfers. I did transfers of fully colonized agar onto fresh agar; two transfers per culture. There were 7 strains in total, so that meant 14 transfers today. The seven strains are south american, brazilian, cambodian, thai lipa yai, hawaiian, texas, and thai mystery. Here's some pictures of the transfers that took place today.









Sorry that there aren't more pictures and they aren't a little higher quality but I'm running short on time to sleep let alone take pictures.


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7614143 - 11/09/07 08:41 AM (16 years, 2 months ago)

I went on a little trip so I haven't up dated in a couple days. The following pictures were taken 11/09/07 I just didn't get a chance to upload them. So, here's some pics of some spawnbags and my agar transfers. We experienced a 25% contamination rate with the spawnbags, but we are fairly sure that this is because there was a shortage of syringes and we were using each individual syringe more times than we probably should have w/o completely re-sterilizing them. We also did not have injection sites and just went straight through the bag and taped in the open air of a far from ideal office. Given these laxed conditions I definitely did not feel bad about the contamination rate. We just got an inoculation gun which will solve our whole syringe contamination vector thing.














The fifth and seventh spawn bags from the top are contamed. On a brighter note my first experiences w/ agar are going fairly well. Out of 9 multispore inoculations and 14 transfers I only experienced two contaminations. Although I did experience these contaminations I'm pretty sure that in some cases the spores were actually no good as I had used a couple really old syringes that had been severely beat to try and hold on to a couple strains but everything knocked up with them contamed, this was the case w/ the ban hua thanon. This is absolutely nothing against the strain it's just the syringe that I used for it had been beat over the last couple years and more than a couple moves.


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InvisibleGlacier Creek
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7617231 - 11/09/07 11:37 PM (16 years, 2 months ago)

nice documentation, thanks!


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Offlinecaptainbonkers
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Re: Getting on it- a Halloween adventure [Re: Glacier Creek]
    #7617299 - 11/10/07 12:07 AM (16 years, 2 months ago)

wow looks lke you're having a pretty thorough go at it psilopod.

curious to see how those spawn bags turn out. i've got a couple on the go myself.

keep the pics coming.


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: Glacier Creek]
    #7617791 - 11/10/07 07:43 AM (16 years, 2 months ago)

I'm trying my best to stay on top of things, but it's hard.  If I'm here updating, I'm not taking care of everything that needs to be done; If I take care of everything that needs to get done, I'm not updating the thread.  I'm balancing it out I guess; I'll try and keep up with the once a day and try to get a little more thorough.  This is supposed to be agar selection isn't it.  I guess I should give some details.

The first thing that comes to mind is that I poured the agar a little thick and the transfer pieces are way bigger than they would ever need to be(thickness).  So, in the future I will definitely be pouring the agar on the thin side that way I get more agar cultures out of it and it's a little less work for the myc as they would't have to colonize the rest of the transfer before getting to the fresh agar.  I suppose putting the transfer in upside down could solve this problem.  I'm not sure whether the lack of light or air would have adverse effects on the transfer though.  If anyone could chime in on this matter it would be appreciated, whether it be about actual pouring and transfer technique, or about the the matter of putting the transfer in upside down to hasten the time it take to make contact with the fresh agar.

So far, of the multispore inoculations(WBS spawnbags/jars) the Brazilian is the absolute fastest hands down.  The only other strain that is remotely close in colonization time in this experiment here so far is South American, which still does not perform on the level of the Brazilian. As far as the agar transfers go Texas is the standout candidate so far.  Hawaiian, South American, Puerto Rican, and Cambodian are not too far behind but they aren't as far as Texas.  The Thai Mystery is the only one that seems to be behind everyone else as far as agar goes.  With the g2g pint jars both the Brazilian and the South American are almost fully colonized.  I don't know how many days that is exactly but that is a pretty fast colonization time.  The amazing thing is that the Brazilian spawnbags are going to be fully colonized very soon as well.  I must say that this mycelium water technique that I've been using for inoculation is beautiful.  You don't have to wait for LC to colonize; you can essentially control the levels of mycelium present in the water; it's instantaneous so long as you have at the very least a partially colonized jar.  Coupling this mycelium water technique with that new inoculation gun should prove beneficial in overall results and happiness levels, lol  :bongload: One thing that was noticed by my friend is that with the strength of the spring in the gun and the repetition of use, our hands should become fairly strong, lol.  The gun only shoots 10ml at a time and we're prolly gonna be hitting stuff with anywhere from 30ml-80ml.  We would inoculate with more(maybe even will) but it depends on the amount of inoculant available at the time. It shouldn't be too much of a problem though.

The last round of spawnbags that we did we decided to bulk up in size.  The last ones were probably about a third full, while this batch is probably a little bit less than half full.  I want to see how far the limits can be pushed per bag, I read what RR posted on this matter but I figured I'd give it a try myself.

On a similar note has anyone had experience inoculating spawnbags of coir with mycelium culture of some sort(lc, water)?  I read about someone experimenting w/ these techniques to hasten time but I could not find anyone that had documented results.  If this was a reasonable technize I would imagine that the preparation of spawn would become exponentially easier, or so I hope :grin:

I would also like to hear everyones opinions on bulk spawn preparation.  I'd like to focus on prep. time/difficulty and as well as any other inherent pros/cons involved in the various possible spawn media(practically endless).

Anyways, I'm gonna go get my day started and actually go check on everything.  Maybe I'll snap a few shots for you guys as well, I'm fairly impressed with the g2g as well as many of the spawnbags performance.


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This and all posts are the work of an imaginary character known as psilopod. Don't believe this rambling drivel, none of it's real, it's fake, it's an illusion, strictly for entertainment purposes. With that being said, enjoy.


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: Glacier Creek]
    #7617798 - 11/10/07 07:46 AM (16 years, 2 months ago)

What exactly do you have mounted in the holes of your glove box it looks like a plastic of some sort. Is that a fan mounted for positive pressure? How's it's overall performance/contam. rate?


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7617906 - 11/10/07 08:50 AM (16 years, 2 months ago)

So here's a little morning update.  Everything seems to be going well, the braz g2g is just about done, the South American g2g is almost there as well.  Quite a few of the bags are well developed as well.  The first pic is the braz g2g and the second pic is the SA g2g.

I've also noticed that the drier BRF that's closest to the heatbomb seems to be the fastest overall runner.  I think that it's due to the low moisture content of the BRF because wetter/clumpier BRF didn't work anywhere near as well.  That's more my buddy's project than mine though, I like grains.  If the BRF is fastest, easy, and works, I'm all about it though.  Anyways, here's some pics from this morning.
Brazilian g2g

South American g2g

Brazilian tearing it up!!! :rockon:













The third agar culture is my favorite so far, it's Texas.  I'll get this edited soon and add labels for the pictures so you guys know which strain is which for the agar.

Brazilian agar transfer



Texas











Cambodian agar transfer



Puerto Rican agar transfer


Edited by psilopod (11/11/07 09:50 AM)


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7618206 - 11/10/07 10:48 AM (16 years, 2 months ago)

We just got a new pressure cooker in the mail from a friend via kickdown:stoned: We now have 2 working Presto 23 qt pressure cookers.  We have one other one as well but it got bent in an accident and isn't exactly perfect to say the least.  Maybe with a little force we can get it back in working condition.  Either way though, 2 23qt pc's will be awesome.


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This and all posts are the work of an imaginary character known as psilopod. Don't believe this rambling drivel, none of it's real, it's fake, it's an illusion, strictly for entertainment purposes. With that being said, enjoy.


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Edited by psilopod (11/10/07 06:12 PM)


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7619873 - 11/10/07 06:18 PM (16 years, 2 months ago)

The inoculation gun is quite the little tool. We haven't used it yet but we tested it out w/ water and it's awesome. I'll also mention that once there is liquid inside of it the gun is much easier to operate/squeeze. We made one slight modification to the gun which was to make a glove that slides over the back barrel because it's exposed and seemed to be a possible vector of contamination. Maybe it's overkill but figured better safe than sorry.

There's also a fridge in the works as well. We already have a large upright fridge/freezer that we are going to be able to use for another incubator we just need to clean it first as it's pretty funky.

I'll have pics of both soon.

Edit
Before
After


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This and all posts are the work of an imaginary character known as psilopod. Don't believe this rambling drivel, none of it's real, it's fake, it's an illusion, strictly for entertainment purposes. With that being said, enjoy.


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Edited by psilopod (11/10/07 08:13 PM)


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7621370 - 11/11/07 09:30 AM (16 years, 2 months ago)

More pics for you guys. I'm trying to be a little bit more thorough in my whole procedure here so I'll try my best to give you guys some detail.  I might have messed some of the code up for the pics so I might have to mess with it a little to make it accurate.

Everything is going at extreme rates.  This is due to an accidental increase in incubator temp. that took place bringing it someone in the near 90 degree area.  We suffered three more contaminations of a bags that were touching the giant heatbomb.  The contamination occured directly where the bag was touching the heatbomb so I know that's what caused it.  Kinda sucks, and we prolly could salvage it, but no need to be messing around w/ funky stuff.  So that's twelve bags in total out of 31 that were contaminated.  I have mixed feelings about this.  In all honesty I feel a little crappy if I focus on it(38.7%contam. rate :shake: )but I do know where we went wrong(identified the vectors of contamination and errors in technique) and we have moved on.  Fortunately we had the foresight to prepare for contamination and did a little extra :bongload: So, although it may hurt the pride a little we are still in good shape. It was basically inevitable that we would mess up but I figured that we would play the percentage game, accomodating for up to 50% contamination within our run and still being in okay shape for our goals.  Not to say that we expected 50% contamination but we knew that there would be alot of little kinks to work out of these little systems especially because we didn't follow anyone else's setup verbatum.  Once we get our whole system dialed in we'll be fine, heck, we're still fine right now; pride's just a little hurt. What can you expect though, we are newbs.  I think that we have all the tools necessary to do just about anything you see on these forums, we just don't have the experience yet.  This is the other reason the I changed the threads title over to the beginning.  Like I said though, we're still doing alright

I'm actually thinking about hooking a pump/hose up to the heatbomb and doing a little radiant heating throughout the freezer.  That way we could turn the heat down a little bit and keep it more controlled/even.

As far as the grain goes Brazilian is hands down the winner so far.  It is basically done colonizing the pint g2g that I did as well as almost being done in some of the spawnbags.  :bongload:

On a more positive note let's hit up some of the new myco porn.  Maybe I shouldn't call it new though, it's from last night. 

Brazilian g2g 1      The g2g's have 2 shots each.  There's only 3 g2g pints in total.

Brazilian g2g 2

Thailand Lipa Yai 1

Thailand Lipa Yai 2

South American 1

South American 2

South American agar transfer 1

Brazilian agar transfer 1

Brazilian agar transfer 2

Cambodian agar transfer 1

Cambodian agar transfer 2

Hawaiian agar transfer 1

Hawaiian agar transfer 2

Puerto Rican agar transfer 1

Puerto Rican agar transfer 2

Thai Mystery agar transfer 1 (looking mighty dank)

Thai Mystery agar transfer 2

Texas agar transfer 1

Texas agar transfer 2

Group shot of first round transfers, two of each strain, in order of my liking so far from left to right: South American, Texas, Hawaiian, Brazilian, Cambodian, Thai Mystery, Puerto Rican.

On the initial multispore inoculation of the agar Puerto Rican was looking like a beast and now it just doesn't seem to have that same flare after being transfered.  I don't know if it's a change in conditions or if it was because of the section I isolated.  I don't know.

With this first round of transfers as mentioned before I am ranking South American as the #1 culture and Texas as the #2 culture, but that's just for now, things could change. :bongload:  :bongload:


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This and all posts are the work of an imaginary character known as psilopod. Don't believe this rambling drivel, none of it's real, it's fake, it's an illusion, strictly for entertainment purposes. With that being said, enjoy.


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7629108 - 11/13/07 07:07 AM (16 years, 2 months ago)

Well, all I can say is BRAZILIAN!! :rockon: It is literally tearing through everything that it touches.  I've actually noticed that the Brazilian almost seems to be isolating itself.  The rhizomorphic growth is so fast that it just dominates.  The rhizomorphic growth is not uniform nor singular but it is definitely visible throughout the various spawns that have been started.







The agar transfers are just about ready for their second round of transfers.  It's hard to tell because I can't see into the top,  but I don't think that I have that much sectoring going on.  The majority of the growth looks like a white mass, not rhizomorphic.  There is rhizomorphic growth though on the edge of the culture that start to shoot up the walls of the jar.  Oh well, maybe it'll take me two more transfers instead of one, who knows. Any advice would be appreciated.
Cambodian agar transfer 11/12/07

Hawaiian agar transfer 11/12/07

Puerto Rican agar transfer 11/12/07

South American agar transfer 11/12/07

Thai Mystery agar transfer 11/12/07

Texas agar transfer 11/12/07

My order of preference for the transfers is currently as follows from well to moderately liked: South American, Hawaiian, Texas, Cambodian, Brazilian, Thai Mystery, Puerto Rican

One more thing that I would like to say, the inoculation gun rocks, it's amazing.  For any of you that have been thinking about investing in one of these things go for it.  If you think it may be remotely practical for you get it, it cuts inoculation time down so drastically.  So, if anyone is looking for a review, I give the inoculation gun 4stars.  It could have had 5 stars had we sprung for the more expensive version, but this one works just fine for $100 less.


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This and all posts are the work of an imaginary character known as psilopod. Don't believe this rambling drivel, none of it's real, it's fake, it's an illusion, strictly for entertainment purposes. With that being said, enjoy.


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Edited by psilopod (11/13/07 07:19 AM)


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Invisiblejeetered
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7629137 - 11/13/07 07:18 AM (16 years, 2 months ago)

those grain bags look heavily tomentose, i see no ropey rhizo whatsoever.


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Offlinepsilopod
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Re: Getting on it- a Halloween adventure [Re: jeetered]
    #7629877 - 11/13/07 11:09 AM (16 years, 2 months ago)

They definitely are tomentose, they were inoculated using mycelium water from a multispore inoculation. This culture has not been worked at all nor has there been any effort made to isolate that particular culture. What I was trying to say and maybe didn't quite say so clearly was the brazilian spawn bags have areas popping up all over the place of pronounced rhizomorphic growth. This is not that significant at this point but it gives me hopes for the future. The pics that I last posted weren't really trying to focus in on the rhizo areas. I will say though that the rhizo area aren't that big and are sporadic but of all the spawn bags going this strain seems to be displaying some of the most desirable characteristics. Again, I know that all of these strains still have to be worked quite a bit, yet another reason why I retitled the original post The Beginning- substrain selection. Either way though, the basic jist of what I'm trying to say is that I'm pleased w/ what is going on right now. Many of my spawnbags are basically fully colonized at this point, 13 days after inoculation. Everything can and does change though; we'll have to see what the future holds.


--------------------
This and all posts are the work of an imaginary character known as psilopod. Don't believe this rambling drivel, none of it's real, it's fake, it's an illusion, strictly for entertainment purposes. With that being said, enjoy.


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Invisiblejeetered
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7630391 - 11/13/07 12:58 PM (16 years, 2 months ago)

could one take a cutting of some very rhizo myc from a section of colonized poo per say? and place it on agar? and clone it that way?
I would assume many transfers would be in play.


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Re: Getting on it- a Halloween adventure [Re: jeetered]
    #7630523 - 11/13/07 01:18 PM (16 years, 2 months ago)

You could definitely slice off a little hunk of the rhizo and transfer it to agar. From there I would imagine that depending on how carefully you sliced you could prolly have it cleaned up in two transfers. Now this is just a guess as I have not done it but I figured it may give some others a little more perspective.

That's basically what I have going on right now, substrain selection. I've got my eye on different traits and then from there I'm just cutting 'em down. I have 14 first generation transfers from multispore inoculations right now on agar. Those transfers will be getting transferred soon bringing it to the second gen. transfer. I'm not sure what the actual terminology is for these procedures but I think you get what I'm saying.

Honestly after these bags fruit out I'll prolly clone a couple of the fruits as well. We'll see how things go. I'm just enjoying myself.

So far I love the brazilians performance on the grain. South American has been the tried and true performer through below ideal conditions. Hawaiian has very strong growth and I'm very interested in it's bioassay.


--------------------
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Re: Getting on it- a Halloween adventure [Re: psilopod]
    #7634411 - 11/14/07 10:51 AM (16 years, 2 months ago)

I have a quick question to throw out there for everybody concerning wood as a substrate. We have recently gained access to a near unlimited supply of hardwood shavings. My question is about wood preparation as a substrate. I read another members postings mentioning a fermented wood additive in substrate, and I'm wondering if something like this should be done or if they could be used as is after being pasteurized. Anyways, if anyone has any experience supplementing cube's substrate with wood a few pointers would be appreciated. Right now we have a large shopping bag of maple wood shavings that are about 3-4" in length, a mm in thickness, and a cm wide. There's a slight possibility that there is some beach wood in the mix as well, but not much.


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This and all posts are the work of an imaginary character known as psilopod. Don't believe this rambling drivel, none of it's real, it's fake, it's an illusion, strictly for entertainment purposes. With that being said, enjoy.


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