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DottoreWolfe
Doctor Wolfe


Registered: 02/21/15
Posts: 201
Last seen: 5 months, 12 days
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Did i ruin my LC jars?
#22079164 - 08/11/15 10:51 PM (8 years, 5 months ago) |
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Greetings guys. So I was preparing some jars for LC and had finished PCing. Just now I looked inside the jars and saw that I must not have cleaned the jars thoroughly, can I still use these or should I redo it. Theres like a few random brown (I think cardboard, because most of these are unused jars, but could be something else) small clumps (comparable to size of spores in a syringe).
-------------------- Everything I post should be regarded as wholly fictitious or hypothetical, nothing I post has any basis in reality.
Edited by DottoreWolfe (08/11/15 10:57 PM)
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Psilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
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If you sterilized the jar long enough, you shouldn't have a problem with some random particles in the water. It might make it harder to tell what is a contam and what is not, once the mycelium starts colonizing the LC, but you should be fine either way, just look carefully under a bright light. Then transfer the healthy mycelium out of the jar.
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DottoreWolfe
Doctor Wolfe


Registered: 02/21/15
Posts: 201
Last seen: 5 months, 12 days
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Quote:
Psilosoulful said: If you sterilized the jar long enough, you shouldn't have a problem with some random particles in the water. It might make it harder to tell what is a contam and what is not, once the mycelium starts colonizing the LC, but you should be fine either way, just look carefully under a bright light. Then transfer the healthy mycelium out of the jar.
This is my first attempt ever using a PC and making LC I PCd for 35ish minutes with 15psi
-------------------- Everything I post should be regarded as wholly fictitious or hypothetical, nothing I post has any basis in reality.
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Psilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
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Yep, that's more than enough time to sterilize a LC jar, you're good to go!
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GerG
Ghost



Registered: 05/27/11
Posts: 50
Loc: From Paris With Love
Last seen: 6 years, 8 months
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Careful with LC. That shit can look great, but end up being totally useless. Make sure to do a test jar or two before you start noc'ing up multiple jars.
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"All you know about me is what I sold you." - Maynard James Keenan
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Psilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
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Re: Did i ruin my LC jars? [Re: GerG]
#22079234 - 08/11/15 11:07 PM (8 years, 5 months ago) |
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Better yet, start your LC with a piece of clean agar next time, I'm assuming you used MS..
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DottoreWolfe
Doctor Wolfe


Registered: 02/21/15
Posts: 201
Last seen: 5 months, 12 days
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Quote:
Psilosoulful said:
Better yet, start your LC with a piece of clean agar next time, I'm assuming you used MS..
Nope, doing a spore print straight to LC, because the vendor only had the print of the exotic strain. http://www.shroomery.org/forums/showflat.php/Number/5238137 The beginner friendly method. Fingers crossed though.
-------------------- Everything I post should be regarded as wholly fictitious or hypothetical, nothing I post has any basis in reality.
Edited by DottoreWolfe (08/12/15 01:34 AM)
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Psilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
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Quote:
DottoreWolfe said:
Quote:
Psilosoulful said:
Better yet, start your LC with a piece of clean agar next time, I'm assuming you used MS..
Nope, I wish, I'm being stupid and doing a spore print straight to LC, because the vendor only had the print of the exotic strain. http://www.shroomery.org/forums/showflat.php/Number/5238137 The beginner friendly method. Fingers crossed though.
Spore prints are never completely contam free, since the fruits are grown in open air. And transferring that to an LC jar, only gives the contams a chance to thrive in a nutrient rich environment. However, there are some growers that have pulled it off and gotten healthy contam free mycelium, but the chances are still pretty low. If you can, I'd suggest getting some agar supplies, following pasty's agar tek, then testing the mycelium from your LC out on the agar dishes FIRST, before going to grains.
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DottoreWolfe
Doctor Wolfe


Registered: 02/21/15
Posts: 201
Last seen: 5 months, 12 days
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Quote:
Psilosoulful said:
Quote:
DottoreWolfe said:
Quote:
Psilosoulful said:
Better yet, start your LC with a piece of clean agar next time, I'm assuming you used MS..
Nope, I wish, I'm being stupid and doing a spore print straight to LC, because the vendor only had the print of the exotic strain. http://www.shroomery.org/forums/showflat.php/Number/5238137 The beginner friendly method. Fingers crossed though.
Spore prints are never completely contam free, since the fruits are grown in open air. And transferring that to an LC jar, only gives the contams a chance to thrive in a nutrient rich environment. However, there are some growers that have pulled it off and gotten healthy contam free mycelium, but the chances are still pretty low. If you can, I'd suggest getting some agar supplies, following pasty's agar tek, then testing the mycelium from your LC out on the agar dishes FIRST, before going to grains.
I don't get how the chances good be so low if in the tek this guy successfully does it everytime, I mean isn't it basically the exact same as using MS into a LC?
-------------------- Everything I post should be regarded as wholly fictitious or hypothetical, nothing I post has any basis in reality.
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maddchef
Vaginal escape artist



Registered: 09/04/09
Posts: 5,602
Loc: Your mom's vag
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On prints its basically more concentrated. You have x amount of contams per centimeter.
If you used square centimeter and dilute it in a 10cc syringe you've spread the contams few and far.
So when you use a drop or two you have a good chance of not getting a contam in the drop
If you scrape that same square directly into an lc then if there was even 1 contam your lc is fucked.
Make sense?
-------------------- In the land of the blind, the one eyed man is king. All mushrooms are edible, but some only once..... Easier than cakes I do science and shit.
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DottoreWolfe
Doctor Wolfe


Registered: 02/21/15
Posts: 201
Last seen: 5 months, 12 days
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Re: Did i ruin my LC jars? [Re: maddchef]
#22079328 - 08/11/15 11:34 PM (8 years, 5 months ago) |
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Quote:
maddchef said: On prints its basically more concentrated. You have x amount of contams per centimeter.
If you used square centimeter and dilute it in a 10cc syringe you've spread the contams few and far.
So when you use a drop or two you have a good chance of not getting a contam in the drop
If you scrape that same square directly into an lc then if there was even 1 contam your lc is fucked.
Make sense?
Fuck...did I just waste 7 LC jars. Or is there actually a solid chance of one or two surviving.
-------------------- Everything I post should be regarded as wholly fictitious or hypothetical, nothing I post has any basis in reality.
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GerG
Ghost



Registered: 05/27/11
Posts: 50
Loc: From Paris With Love
Last seen: 6 years, 8 months
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Re: Did i ruin my LC jars? [Re: maddchef]
#22079366 - 08/11/15 11:43 PM (8 years, 5 months ago) |
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Quote:
I don't get how the chances good be so low if in the tek this guy successfully does it everytime, I mean isn't it basically the exact same as using MS into a LC?
Absolutely nothing works EVERY time. Everything, every method, every tek will fail at some point.
I hate to admit it, but I use LC alot. Even though I am as sterile as possible during every step, I still only have around a 75-85% success rate with LC. And I still don't believe that is an acceptable success rate. Like I said; test everything before using it on anything (except said test jar).
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"All you know about me is what I sold you." - Maynard James Keenan
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Psilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
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You may not see it in your LC jar once the mycelium starts forming, that's why I suggested testing it out on an agar dish because any contams present WILL show up!! Agar is easy to make and is a necessary tool in getting a healthy culture to work with.
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DottoreWolfe
Doctor Wolfe


Registered: 02/21/15
Posts: 201
Last seen: 5 months, 12 days
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Quote:
Psilosoulful said: You may not see it in your LC jar once the mycelium starts forming, that's why I suggested testing it out on an agar dish because any contams present WILL show up!! Agar is easy to make and is a necessary tool in getting a healthy culture to work with. 
Instead of testing in agar can I just inoculate one rye grain jar with LC from each of my LC jars. Then wait to see which jars develop contams and then just not use those LC jars?
-------------------- Everything I post should be regarded as wholly fictitious or hypothetical, nothing I post has any basis in reality.
Edited by DottoreWolfe (08/12/15 12:20 AM)
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GerG
Ghost



Registered: 05/27/11
Posts: 50
Loc: From Paris With Love
Last seen: 6 years, 8 months
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Quote:
Instead of testing in agar can I just inoculate one rye grain jar with LC from each of my LC jars. Then wait to see which jars develop contams and then just not use those LC jars?
Yes.
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"All you know about me is what I sold you." - Maynard James Keenan
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Quote:
DottoreWolfe said:
Quote:
Psilosoulful said: You may not see it in your LC jar once the mycelium starts forming, that's why I suggested testing it out on an agar dish because any contams present WILL show up!! Agar is easy to make and is a necessary tool in getting a healthy culture to work with. 
Instead of testing in agar can I just inoculate one rye grain jar with LC from each of my LC jars. Then wait to see which jars develop contams and then just not use those LC jars?
Testing is testing
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mossystone14
Stranger
Registered: 08/12/15
Posts: 54
Last seen: 8 years, 5 months
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Re: Did i ruin my LC jars? *DELETED* [Re: bodhisatta]
#22080101 - 08/12/15 06:35 AM (8 years, 5 months ago) |
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Post deleted by mossystone14Reason for deletion: my bad
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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That's because inexperienced cultivators think it is a magic bullet that will save them time or make it easier. It's like giving someone a 1000cc crotch rocket for their 16th birthday. A few kids won't kill themselves but most will.
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Psilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
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Quote:
DottoreWolfe said:
Quote:
Psilosoulful said: You may not see it in your LC jar once the mycelium starts forming, that's why I suggested testing it out on an agar dish because any contams present WILL show up!! Agar is easy to make and is a necessary tool in getting a healthy culture to work with. 
Instead of testing in agar can I just inoculate one rye grain jar with LC from each of my LC jars. Then wait to see which jars develop contams and then just not use those LC jars?
Honestly, I'd rather test my LC on a small agar dish (less PC time, quicker results) than having to go through the process of testing it on grain jars. Another reason not to go from LC>grain, is that contams can easily hide within a grain jar and not be noticed, whereas contams have nowhere to hide on agar.
Do you really want to spawn grain jars to bulk, only to find out later that they were actually "dirty" spawn jars and your first flush was sub-par or you got trich after full colonization.
There is a reason why agar is HIGHLY recommended by almost every trusted cultivator on this site as a starting point to test your LC's, GLC's, MS, etc...
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DottoreWolfe
Doctor Wolfe


Registered: 02/21/15
Posts: 201
Last seen: 5 months, 12 days
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Darn it! I don't have any agar on hand and money is kind of tight. Is agar agar, water and a pertri dish all I need to test with the agar?
-------------------- Everything I post should be regarded as wholly fictitious or hypothetical, nothing I post has any basis in reality.
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Psilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
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You need agar agar (I got mine from ebay), glad mini rounds, potato flakes, some light malt karo or honey, micropore tape, and a PC. Check out Pasty's tek, that's where most people start when they dip their toes into agar.  http://www.shroomery.org/forums/showflat.php/Number/19208976
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shellzenone
Zen



Registered: 12/11/14
Posts: 688
Last seen: 1 year, 8 months
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Just wondering OP, did you use metal lids? I have found that sometimes rust particles end up dropping down to the LC. Im thinking about switching to plastic lids for this reason.
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DottoreWolfe
Doctor Wolfe


Registered: 02/21/15
Posts: 201
Last seen: 5 months, 12 days
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Quote:
Psilosoulful said: You need agar agar (I got mine from ebay), glad mini rounds, potato flakes, some light malt karo or honey, micropore tape, and a PC. Check out Pasty's tek, that's where most people start when they dip their toes into agar.  http://www.shroomery.org/forums/showflat.php/Number/19208976
I was doing research all morning, but I found a buddy who just happened to have some shroomsupply MaltExtract Agar lying around, can I use this with petri dishes (the first recipe on here is for same thing http://www.shroomery.org/5243/What-are-some-good-agar-media-recipes). How would I use this? It says it needs to be sterilized, do I sterilize with water added after or water added before sterilization? Do I sterilize it in a quart jar and then transfer to petri dishes or do I put in petris during PC?
-------------------- Everything I post should be regarded as wholly fictitious or hypothetical, nothing I post has any basis in reality.
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Psilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
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I've never done the poured agar tek before, but you are supposed to sterilize your agar mix in a glass beaker with tin foil (loosely) over the top for about 30 min at 15 psi, then once the PC cools down, take out your jar and when it feels comfortable to hold but not too hot that it burns your hand, pour the agar into your petri dishes (in a SAB of course). Put the lids on the petris and let it cool down in the SAB overnight.
Add the water to your agar recipe beforehand, that's what makes it liquid agar. When it cools down, it will become gel-like and ready to use.
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Juiceh
Dabbing All Day



Registered: 09/25/12
Posts: 3,208
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Don't use a beaker for pouring agar. You want something with a smaller opening. Like an empty liquor bottle. Or a Pyrex media vessel. I mean, you could use a beaker if you want, but it's not the best option. Hell, there are people that do it with qt jars.
Add 500ml of water to a pot and put it on the stove and get it hot, but not boiling. Then slowly add your required volume of premade agar mixture while stirring constantly. If you are using a non premixed agar, add all dry ingredients slowly and stir them in before you begin adding the agar agar. Once all your dry ingredients including the agar agar have been added and dissolved into the pot of hot water, pour that then into your glass agar vessel(liquor bottle) with a funnel.
I have a 1/4" hole drilled into the lid of my cuervo bottle with a sfd patch on top of it, which I then cover with foil. After so many uses the thread on the cap for the cuervo bottle have gone shitty and its pretty loose so the filter doesn't do much. When I use my pyrex media vessels I just leave the cap 1/4 turn loose. If you do not have a flowhood, it is preferable to have a filter of some sort on the agar vessel. As your PC cools it will draw in outside air(unless you have a sterilizer that seals and pulls vacuum) this will get drawn into your agar vessel if there is no filter. Before I upgraded my PCs to sterilizers I would move my PC in front of my flowhood for cool down.
Once the PC has cooled enough and there is no pressure, open it up and take out your agar bottle(I do all this in front of a flow hood). Once the agar has dropped below 130F(checked with an infrared thermometer) I begin pouring the plates in front of the hood. I like to stack the empty petris in stacks of 5 all stacked on top of each other in an offset fashion. When I begin pouring I take 5 off the top of the stack and place it next to the stack and begin pouring into the bottom dish of the 5 by lifting the lid and picking up the stack of 4 above. After every 5 dishes I grab the next 5 and place it on top of the stack of 5 that were just poured and repeat.
Here are some pics:

Note the offset stacking of the unpoured dishes in this pic and the stack of poured dishes:

Agar is hot coming out of the PC, be careful:

It probably takes about 90 seconds to pour a whole sleeve of 20 plates.
Quote:
DottoreWolfe said:
Quote:
Psilosoulful said:
Better yet, start your LC with a piece of clean agar next time, I'm assuming you used MS..
Nope, doing a spore print straight to LC, because the vendor only had the print of the exotic strain. http://www.shroomery.org/forums/showflat.php/Number/5238137 The beginner friendly method. Fingers crossed though.
Spore print inoculations are multispore. And prints to LC are risky, so are syringes to LC. The only LC inoculation I would recommend is an agar wedge, or in the case of sclerotia forming species transfering a stone from agar to LC. That tek you linked is ancient and IMHO very flawed.
Edited by Juiceh (08/12/15 04:18 PM)
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: Did i ruin my LC jars? [Re: Juiceh]
#22082389 - 08/12/15 05:13 PM (8 years, 5 months ago) |
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DottoreWolfe
Doctor Wolfe


Registered: 02/21/15
Posts: 201
Last seen: 5 months, 12 days
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Re: Did i ruin my LC jars? [Re: bodhisatta]
#22083204 - 08/12/15 08:08 PM (8 years, 5 months ago) |
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Thanks a ton guys hopefully at least one of these LCs produces a clean agar.
-------------------- Everything I post should be regarded as wholly fictitious or hypothetical, nothing I post has any basis in reality.
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_OttO_
Over Stimulated



Registered: 06/01/05
Posts: 2,588
Loc: Up Over
Last seen: 1 month, 13 days
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Just be patient, you'll know soon enough. You can test it on a small grain jar, but i never tested any LC because I'm still yet to have one fail...
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