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Violet



Registered: 12/06/11
Posts: 4,205
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Re: Culturing • GRAIN 'PETRI' TEK [Re: BigGreenMat]
#19176850 - 11/22/13 10:48 PM (10 years, 2 months ago) |
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Quote:
BigGreenMat said: Hey Violet, I see you going back and forth between the agar petri and the grain petri, but can you isolate and transfer straight grain petri to grain petri. Can you go from fruit to grain petri? Can you go from spore to grain petri?
Can you go from fruit to grain petri? Yes you can, but I wouldn't recommend it. After a clone you want to isolate the strongest growth from that clone and this is possible on the 2D of straight agar.
Can you go from spore to grain petri? Yes you definitely can, but mostly I wouldn't suggest it, except a certain way. If you have a clean print, and are using SAB (Not laminar airflow for this one), you can drop spores onto the middle area of the grains. Hopefully the top of the grains will be adequate, which it will be if containers are air-tight and the ratio of grain to air not too shallow. Better for this would be spore solution, which is the certain way I mentioned. Grain petris are the perfect and ideal starting medium for spore solution when the intent is not to do a full-size multispore grow. Just 2-3 drops is perfect, I suggest only in the middle just like agar wedge transfer. In this scenario the spore solution has its best chance of germ without bacteria, just the same as with jars, and also the advantage of uni-centric outward growth like on agar - in fact it's the only easy/assured way of getting such growth from spores on agar.
The big question is your first: Can you isolate and transfer grain petri to grain petri?
Firstly, yes you definitely can easily and excellently transfer from grain petris to others. Tweezers are the perfect tool in general, and getting to use them for mycology work instead of scalpel-juggling/scraping is absolutely heaven. With tweezers, a single tiny grain petri is hand-transferred to dozens of containers in minutes with great ease.
Isolation... This I'm not entirely sure about, but my guess is No. The mycelium within the grains grows 3D and we do not see "sectoring" amongst grains with this growth. Though it's likely it would eventually happen with transfers, it would not be possible or easy to isolate genetics in this fashion. That's much more effective and assured on agar.
-------------------- Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it! PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers The simplest, quickest, safest tek! For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers! • Violet's Teks and Posts •
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anne halonium
jaguarette


Registered: 05/07/13
Posts: 1,908
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Re: Culturing Tech • Agar containers, [Re: JMcDoogle]
#19177858 - 11/23/13 06:58 AM (10 years, 2 months ago) |
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pay no attention to JM, lemme spell it out for ya so theres zero doubt.
Quote:
JMcDoogle said: Frank Horrigan - Plenty of References.
now heres a quote from his trusty master.........
Quote:
FrankHorrigan said: You don't even have to be sterile with a colonized grain. I used to make isolates from grains that fell to the floor of my SAB during G2G. Just pick it up when you are done with your work and place it on a clean dish 
^sounds more crusty , than trusty to me.
its no surprise , that these posers defame anything that doesnt involve turds in buckets. they are literally unable to master sterile skills beyond that.
JM pops up on threads, to misinform and slander. and, has nothing to offer a serious grower.
if anything JM, frank is a reference for your grow skill, and his own.
now back to Vtek.........the grow tek so cool, that it drives peeps that lack grow skill , insane.
Edited by anne halonium (11/23/13 07:21 AM)
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JMcDoogle
A Serious Scholar


Registered: 07/07/09
Posts: 1,495
Loc: Nunavut
Last seen: 3 years, 9 months
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Re: Culturing Tech • Agar containers, [Re: anne halonium]
#19177895 - 11/23/13 07:24 AM (10 years, 2 months ago) |
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Quote:
anne halonium said: pay no attention to JM, lemme spell it out for ya so theres zero doubt.
Quote:
JMcDoogle said: Frank Horrigan - Plenty of References.
now heres a quote from his trusty master.........
Quote:
FrankHorrigan said: You don't even have to be sterile with a colonized grain. I used to make isolates from grains that fell to the floor of my SAB during G2G. Just pick it up when you are done with your work and place it on a clean dish 
^sounds more crusty , than trusty to me.
its no surprise , that these posers defame anything that doesnt involve turds in buckets. they are literally unable to master sterile skills beyond that.
JM pops up on threads, to misinform and slander. and, has nothing to offer a serious grower.
if anything JM, frank is a reference for your grow skill, and his own.
now back to Vtek.........the grow tek so cool, that it drives peeps that lack grow skill , insane.
I find this insulting - You're directly aiming insults at my character and I will not have that.
How you like me now.

Oh wait, I dont care what you say Anne, nor does half of the people who come across your posts.
Would you like to know why?
You're bat-shit crazy, thats why.
Thanks.
This tek is a wonderful beginning, I like where its headed. Could use some finer-tuning but hell, we all start somewhere.
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The ego is nothing other than the focus of conscious attention.
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anne halonium
jaguarette


Registered: 05/07/13
Posts: 1,908
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Re: Culturing Tech • Agar containers, [Re: JMcDoogle]
#19177906 - 11/23/13 07:31 AM (10 years, 2 months ago) |
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glad ya quoted it, i would have gladly said it 2 x............
now, on with the show
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BigGreenMat
Stranger

Registered: 11/04/13
Posts: 161
Last seen: 3 years, 3 months
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Re: Culturing • GRAIN 'PETRI' TEK [Re: Violet]
#19179058 - 11/23/13 01:50 PM (10 years, 2 months ago) |
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Quote:
Violet said:
Quote:
BigGreenMat said: Hey Violet, I see you going back and forth between the agar petri and the grain petri, but can you isolate and transfer straight grain petri to grain petri. Can you go from fruit to grain petri? Can you go from spore to grain petri?
Can you go from fruit to grain petri? Yes you can, but I wouldn't recommend it. After a clone you want to isolate the strongest growth from that clone and this is possible on the 2D of straight agar.
Can you go from spore to grain petri? Yes you definitely can, but mostly I wouldn't suggest it, except a certain way.
Just 2-3 drops is perfect, I suggest only in the middle just like agar wedge transfer. In this scenario the spore solution has its best chance of germ without bacteria, just the same as with jars, and also the advantage of uni-centric outward growth like on agar - in fact it's the only easy/assured way of getting such growth from spores on agar.
The big question is your first: Can you isolate and transfer grain petri to grain petri?
Firstly, yes you definitely can easily and excellently transfer from grain petris to others. Tweezers are the perfect tool in general, and getting to use them for mycology work instead of scalpel-juggling/scraping is absolutely heaven. With tweezers, a single tiny grain petri is hand-transferred to dozens of containers in minutes with great ease.
Awesome thanks for the info.
Here is what I am thinking. I have a few different syringes with different varieties. I was thinking to make up a batch of your grain petris and inoculate them from 2 different syringes (I am thinking my Albino PE and Pan Cyan to try it out). Let the petris grow out then use them to transfer to either RGS or Brown Rice according to your V container tek, and then run them out from there to fruiting. After that I would then transfer a fruit to Agar for some isolation work. Does that sound like a reasonable use of your teks?
Thanks again for all your answers.
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MMagg
Stranger
Registered: 11/19/13
Posts: 41
Last seen: 4 years, 8 months
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Re: Culturing • GRAIN 'PETRI' TEK [Re: BigGreenMat]
#19179174 - 11/23/13 02:19 PM (10 years, 2 months ago) |
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Quote:
BigGreenMat said:
Quote:
Violet said:
Quote:
BigGreenMat said: Hey Violet, I see you going back and forth between the agar petri and the grain petri, but can you isolate and transfer straight grain petri to grain petri. Can you go from fruit to grain petri? Can you go from spore to grain petri?
Can you go from fruit to grain petri? Yes you can, but I wouldn't recommend it. After a clone you want to isolate the strongest growth from that clone and this is possible on the 2D of straight agar.
Can you go from spore to grain petri? Yes you definitely can, but mostly I wouldn't suggest it, except a certain way.
Just 2-3 drops is perfect, I suggest only in the middle just like agar wedge transfer. In this scenario the spore solution has its best chance of germ without bacteria, just the same as with jars, and also the advantage of uni-centric outward growth like on agar - in fact it's the only easy/assured way of getting such growth from spores on agar.
The big question is your first: Can you isolate and transfer grain petri to grain petri?
Firstly, yes you definitely can easily and excellently transfer from grain petris to others. Tweezers are the perfect tool in general, and getting to use them for mycology work instead of scalpel-juggling/scraping is absolutely heaven. With tweezers, a single tiny grain petri is hand-transferred to dozens of containers in minutes with great ease.
Awesome thanks for the info.
Here is what I am thinking. I have a few different syringes with different varieties. I was thinking to make up a batch of your grain petris and inoculate them from 2 different syringes (I am thinking my Albino PE and Pan Cyan to try it out). Let the petris grow out then use them to transfer to either RGS or Brown Rice according to your V container tek, and then run them out from there to fruiting. After that I would then transfer a fruit to Agar for some isolation work. Does that sound like a reasonable use of your teks?
Thanks again for all your answers.
I think she recommends going from spore syringe to thin RGS layer in container.... then take a strain using tweezers to agar.... transfer as need to get isolation, then agar wedge back to 1 cup RGS in container for colonization
Seems to prefer that over spore syringe to agar....... just from what I read of her tek.
Edited by MMagg (11/23/13 02:20 PM)
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BigGreenMat
Stranger

Registered: 11/04/13
Posts: 161
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Re: Culturing • GRAIN 'PETRI' TEK [Re: MMagg]
#19179221 - 11/23/13 02:32 PM (10 years, 2 months ago) |
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Quote:
MMagg said:
I think she recommends going from spore syringe to thing grain layer in container.... then take a strain using tweezers to agar.... transfer as need to get isolation, then agar wedge back to grain container.
Seems to prefer that over spore syringe to agar....... just from what I read of her tek.
I see what you are suggesting, but without some sort of fruiting phase you can't choose good genetics. You could be isolating mycelium which will be poor fruiters and you would never even know. So from all my reading the flow always goes: Spore->Mycelium->Pin/Fruit->Agar for isolation.
Are you suggesting that I try to fruit or pin the initial rice petri and move to agar from that? I could see that being a possibility too.
Oh and welcome fellow neophyte MMagg
Edited by BigGreenMat (11/23/13 02:34 PM)
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Violet



Registered: 12/06/11
Posts: 4,205
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Re: Culturing • GRAIN 'PETRI' TEK [Re: BigGreenMat]
#19180095 - 11/23/13 06:50 PM (10 years, 2 months ago) |
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To give you an idea of my experience with and feelings towards full-size grain cakes of multispore... Ugh. Yeah typically I get 9-18 dry from them when (if) I follow through, at least that's not too bad. They're kindof annoying to harvest particularly having to clean off side/under-pins which is quite likely to happen. Mostly single fruits.
My last run if 24 "grain petris" with no duplicate genetics from one variety's multi-spore agar germination on 2 dishes, I only took about 12 total samples and several of those were multiples from single containers. Taking the best sectors and best dishes, they somewhat divided out to total 24 isolate cultures to test. Of those I'm likely to find that some aren't really heavy fruiters, some may be slow fruiting and end up not having enough flushes, some may yield well but still delay too long for flushes, some may have prolific but small fruits... Some marked from the same clone may also be duplicates... So from 24 separate multi-spore inoculations, all with several spore flake germs of their own thus countless genetic mixes, I hope to get 1, maybe 3, really good isolates to put to a final race for time & yield & potency. That is my opinion of the proportion of good genes from the wildcard of multispore.
-------------------- Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it! PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers The simplest, quickest, safest tek! For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers! • Violet's Teks and Posts •
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blindingleaf
blue collar underworld



Registered: 07/19/13
Posts: 22,008
Loc: sub-surface unseen
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Re: Culturing • GRAIN 'PETRI' TEK [Re: Violet]
#19180140 - 11/23/13 07:06 PM (10 years, 2 months ago) |
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Quote:
Violet said:
My last run if 24 "grain petris" with no duplicate genetics from one variety's multi-spore agar germination on 2 dishes, I only took about 12 total samples and several of those were multiples from single containers. Taking the best sectors and best dishes, they somewhat divided out to total 24 isolate cultures to test. Of those I'm likely to find that some aren't really heavy fruiters, some may be slow fruiting and end up not having enough flushes, some may yield well but still delay too long for flushes, some may have prolific but small fruits... Some marked from the same clone may also be duplicates... So from 24 separate multi-spore inoculations, all with several spore flake germs of their own thus countless genetic mixes, I hope to get 1, maybe 3, really good isolates to put to a final race for time & yield & potency. That is my opinion of the proportion of good genes from the wildcard of multispore.
Hey violet! can u clarify for me this part? when u say u took 12 total samples, I assume u mean from the ms petri, correct? were these fruit samples or agar wedge samples? those 12 sample grew out, and then from there u transferred the 24 most rhyzo mycelium? and they ended up being isolates at that point by luck? or out of those 12 initial samples, 24 isolates were produced after numerous transfers? sorry if I misread and this is a dumb question. Im confused and intrigued, because this is what I want to get into next, just got my PP5's.
-------------------- A few thoughts on cultivation MICROBIAL HUSBANDRY!!!! The whole is greater than the sum of its parts
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Violet



Registered: 12/06/11
Posts: 4,205
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Re: Culturing • GRAIN 'PETRI' TEK [Re: blindingleaf]
#19180200 - 11/23/13 07:27 PM (10 years, 2 months ago) |
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Yeah I took 12 pins total (from maybe only 4-8 of the 24 containers) to 12 agar dishes, and from there took transfers from most of those dishes, which either right away or after 2-3 transfers went thru sectoring where I took the best growth. In this case I came out to 24 dishes of isolated genetics. I'm pretty picky for strong white growth and favor thick ropey more than cottony, although sometimes a culture switches for various reasons.
That's quite a large culture round I did, but it's done in the hopes of assuredly ending up with having found some wonderful strains.
Transferring to another requires at least 2 dishes at the same time, a sending and at least one receiving. For the beginner who hasn't amassed enough of a collection to be able to bounce from that many containers to just as many more, surely not as many cultures can come of a round. Over time compare the winning cultures of whatever size culturing round you can transfer containers for.
-------------------- Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it! PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers The simplest, quickest, safest tek! For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers! • Violet's Teks and Posts •
Edited by Violet (11/23/13 11:26 PM)
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BigGreenMat
Stranger

Registered: 11/04/13
Posts: 161
Last seen: 3 years, 3 months
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Re: Culturing • GRAIN 'PETRI' TEK [Re: Violet]
#19180840 - 11/23/13 11:17 PM (10 years, 2 months ago) |
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Ok, yeah I won't be doing anywhere near that volume. I was thinking of starting on the rice petri and going to agar after a fruit so I could pick something decent to start with. I can see why, at the volumes you talk about, you wouldn't bother even doing a multispore container. I guess I kind of just have to jump in and do it and then see how it pans out and hopefully get some good fruits to clone and isolate.
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Mosey3012



Registered: 12/12/12
Posts: 675
Last seen: 3 months, 15 days
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Re: Culturing • GRAIN 'PETRI' TEK [Re: BigGreenMat]
#19187294 - 11/25/13 02:27 PM (10 years, 2 months ago) |
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Well out of all my plates 1 did turn out to show some growth, and in a surprisingly quick period once it started I can only assume. I feel like I check these just about daily but I must admit I must have been overlooking this plate... Plate marked 11-12-13, growth is about the size quarter. Definitely no sign of life within first 7 days after making clone xfer. Checked on it this morning and found this... So the grain water does work! Even just taking the left over soak water and throwing in .5 agar per 25ml
Close Up

Out Some

Unrelated to the topic but in relation to the the provided pictures... Notice how fluffy that has grown out? Is this normal for clones or in general to grow out that fluffy? I can only assume there's no visible ropeyness from any rizo growth because there's so many different sets of genetics growing out so close to each other? All of my plates seem to grow out this way. Now in these pics (surprisingly) you're able to see on the outter edge of the growing ring some sectoring occurring. I marked off what to my knowledge in the green would more then likely be tomentose growth then in blue where I'd likely get more ropeyness if making a transfer from there? *Zoom in pic to see markings* (We can call left green sector A, Blue: B, and to the right green sector C)

I appreciate any feedback on that haha, I'd like to one day have some iso's under my belt!
-------------------- "Each of these lives is the right one! Every path is the right path. Everything could have been anything else and it would have just as much meaning"
  "All that is gold does not glitter, Not all those who wander are lost"
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WillSolvem
Odd-Hand



Registered: 04/24/12
Posts: 1,519
Loc: Chapter 26
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Re: Culturing • GRAIN 'PETRI' TEK [Re: Mosey3012]
#19187977 - 11/25/13 05:25 PM (10 years, 2 months ago) |
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I'm going to use SFD's for sterile fae, but you really sold me in the PP5 containers 
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AMU Q&A Thread because questions deserve answers.
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WillSolvem
Odd-Hand



Registered: 04/24/12
Posts: 1,519
Loc: Chapter 26
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Re: Culturing • GRAIN 'PETRI' TEK [Re: WillSolvem]
#19189117 - 11/25/13 09:17 PM (10 years, 2 months ago) |
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So here's where I'm at so far:
Using "Friendly Felt" I made some filter discs


Then I abraded around the center so the RTV would adhere well, then drilled a 1/4" hole in the center

I double up the friendly felt, if you notice this stuff has a "rough" side and a "smooth" side. I apply a bead of RTV around the rim on the "smooth" side

And firmly apply to lid

Then lay a bead of RTV around the rim of the "rough" side of the disc

Then I drop another filter disc onto this one, "rough" side down so the "rough" sides of the two filter discs are facing each other and only the "smooth" side will be exposed when finished

Then press firmly, some RTV will push out of the sides, just run your finger around the discs to smooth out the excess

I stuck them in my super high tech anti-cat-curiosity-chamber for the night

I'll update tomorrow after I fill, sterilize, and inoculate. I'll keep posting as the isolates I obtain are expanded and fruited. Thank you Violet for the inspiration and beautiful write-up
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AMU Q&A Thread because questions deserve answers.
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MMagg
Stranger

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Re: Culturing • GRAIN 'PETRI' TEK [Re: WillSolvem]
#19189617 - 11/25/13 11:01 PM (10 years, 2 months ago) |
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Will,
Just curious.... why did you choose to modify the lid when Violet repeatedly advises not to do so? I've got 20 containers cooling off for noc myself.... I didn't modify the lids...
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WillSolvem
Odd-Hand



Registered: 04/24/12
Posts: 1,519
Loc: Chapter 26
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Re: Culturing • GRAIN 'PETRI' TEK [Re: MMagg]
#19190536 - 11/26/13 05:37 AM (10 years, 2 months ago) |
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Quote:
MMagg said: Will,
Just curious.... why did you choose to modify the lid when Violet repeatedly advises not to do so? I've got 20 containers cooling off for noc myself.... I didn't modify the lids...
That's how I have been doing this long before this Tek ever existed
Proof in pudding: http://www.shroomery.org/forums/showflat.php/Number/17459279/page/2
The following is the post I made briefly outlining the technique from the above link
Quote:
Check this out:

Pinning: ksss strain
I fill a jar about 1/4 the way with hpoo/strawn/grain/gypsum, and case with verm, sterilize and inoculate (spores will make it through the verm layer) and wait until colonization/consolidation/fruiting, take clones with a syringe through a hole drilled though the jar and sealed with rtv, use a large hole and a thick layer of rtv so you can move the needle around with confidence, open under your flow hood/SAB for a sterile mushroom for printing or further cloning
It can even be done using the PF tek.. 
 Quote:
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AMU Q&A Thread because questions deserve answers.
Edited by WillSolvem (11/26/13 05:48 AM)
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MMagg
Stranger

Registered: 11/19/13
Posts: 41
Last seen: 4 years, 8 months
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Re: Culturing • GRAIN 'PETRI' TEK [Re: WillSolvem]
#19190992 - 11/26/13 09:18 AM (10 years, 2 months ago) |
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Right, but those are glass jars, not the PP5 plastics.
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anne halonium
jaguarette


Registered: 05/07/13
Posts: 1,908
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Re: Culturing • GRAIN 'PETRI' TEK [Re: MMagg]
#19191006 - 11/26/13 09:25 AM (10 years, 2 months ago) |
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part of the value of PP5 , is they can be stored and stacked discretely, and alot of peeps have collections of storage containers.
its a little stealthier than the stack of masons. modified lids, sorta take away the stealth factor.
we never mod lids, as our experience shows, they get plenty of FAE with loose lids.
some peeps , like to drill and modify and build. i dont see any problem with that.
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MMagg
Stranger

Registered: 11/19/13
Posts: 41
Last seen: 4 years, 8 months
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Quote:
anne halonium said: part of the value of PP5 , is they can be stored and stacked discretely, and alot of peeps have collections of storage containers.
its a little stealthier than the stack of masons. modified lids, sorta take away the stealth factor.
we never mod lids, as our experience shows, they get plenty of FAE with loose lids.
some peeps , like to drill and modify and build. i dont see any problem with that.
Just curios that Violet said she modified the lids when she started, but said she wished she hadn't. Advised against it, but didn't give much reason other than it was not necessary..... as FAE occurs with a loose lid.
Normally, when someone goes out of their way to advise against something, there is a reason... I just wondered if that was the case here....
Edited by MMagg (11/26/13 12:04 PM)
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anne halonium
jaguarette


Registered: 05/07/13
Posts: 1,908
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Re: Culturing • GRAIN 'PETRI' TEK [Re: MMagg]
#19191692 - 11/26/13 12:05 PM (10 years, 2 months ago) |
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well, ya can do it, or not. some peeps like to mod things.
technically i see no harm. question is , why? especially when ya have alot of em.
alot of the violet tek, is designed to avoid extra things.
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