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OregonChronic
Iron, Lion, Cyans
Registered: 09/12/10
Posts: 1,970
Loc: Scoping Woodchips
Last seen: 11 years, 2 months
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Multiply Spores Tek for n00bs 1
#15432426 - 11/27/11 07:29 PM (12 years, 4 months ago) |
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This 'tek' goes out to all the noobs who honor their one spore syringe because they don't know how to multiply cultures yet. Here is how to turn 2-5cc's of an original spore syringe into many more.
You'll need
- 1 spore syringe (must have lots of visible spores)
- a tube of RTV silicone high heat resistant red kind
-a wide mouth half pint jar
-pre packaged, sterile syringes (I get mine from an animal feed store)
-3M micropore tape and tyvek mail envelopes
-distilled water
-still air box (prefered)
Alright. It's very simple. You prep the lid of the half pint jar by puncturing or drilling a 1/8" or 1/4" size inoc hole on the very side without messing up the seal. Then make a bit bigger of a hole on the exact other side of the lid. If you don't have a drill gun you can puncture a bunch of small nail holes.
After you get the holes done, blob a bit of the RTV silicone over the small hole on the side of the bottom of the lid. So it kinda 'sinks' through the hole to the top. Make sure it has a good 'seal' and won't leak or anything. Let harden for 24-72 hours until no more silicone smell.
Then cut a square piece of tyvek from a tyvek envelope and seal it over the top of the big hole with the micropore tape.
The lid should look something like the pics above. It's better to have the tyvek square a bit bigger as it shrinks when sterilized, pulling on the micropore seal. And it might be better to use two layers of micropore for extra thickness though I've had no problems using one layer to seal.
Just fill the clean 1/2pint jar with however much cc's of distilled water and put the lid on tight. Cover with alum foil as usual, and PC for 30mins 15psi on top of a qt jar to raise out the water. If you don't have a PC just steam sterilize for 90-120mins same way.
After it cools down you put the 1/2pint jar in the still air box(optional but preferd) and take the alum foil off inside the box and inoc with 3-5cc's of spore solution through the silicone port. Remember to flame needle. You now have x amount of new sterile spore solution.
Then take the pre packaged sterile syringes and one by one, aspirate the spore solution into each syringe through the silicone port. No need to flame needle since it should be pre packaged and sterile. But you can if you wanna take precautions. You can also use your own syringes if you sterilize them in alum foil before hand.
This can be done in open air but I recommend a still air box,...u never know what can land on that silicone port as soon as the alum foil comes off.
-------------------- Wavy Cap Cyan Spore Prints for Trade/Sale
Edited by OregonChronic (12/02/11 12:15 PM)
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OregonChronic
Iron, Lion, Cyans
Registered: 09/12/10
Posts: 1,970
Loc: Scoping Woodchips
Last seen: 11 years, 2 months
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And make sure the silicone fully cures
You don't want acetic acid getting leeched from the silicone into the sporewater
-------------------- Wavy Cap Cyan Spore Prints for Trade/Sale
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wasabi-kun
easily bored
Registered: 01/24/11
Posts: 169
Loc: where is ware
Last seen: 10 years, 11 months
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nice, should help alot of people
-------------------- ciao-su
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OregonChronic
Iron, Lion, Cyans
Registered: 09/12/10
Posts: 1,970
Loc: Scoping Woodchips
Last seen: 11 years, 2 months
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Re: Multiply Spores Tek for n00bs [Re: wasabi-kun]
#15432588 - 11/27/11 08:03 PM (12 years, 4 months ago) |
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Yea and it helps noobs get more exp for lid prep for grains and over all ideas of how to multiply cultures.
-------------------- Wavy Cap Cyan Spore Prints for Trade/Sale
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iron
I fart loud.
Registered: 11/03/11
Posts: 351
Last seen: 5 years, 4 months
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tripdawg420
low life with no life
Registered: 02/02/09
Posts: 7,076
Loc: illinois
Last seen: 20 hours, 2 minutes
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Re: Multiply Spores Tek for n00bs [Re: iron]
#15432698 - 11/27/11 08:23 PM (12 years, 4 months ago) |
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alright thats pretty cool
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Shroom_Goon
Just Some Guy
Registered: 08/11/11
Posts: 475
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Re: Multiply Spores Tek for n00bs [Re: tripdawg420]
#15432770 - 11/27/11 08:39 PM (12 years, 4 months ago) |
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Simple enough
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hawkington
Registered: 11/23/11
Posts: 29
Last seen: 12 years, 3 months
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Re: Multiply Spores Tek for n00bs [Re: Shroom_Goon]
#15454037 - 12/02/11 10:27 AM (12 years, 4 months ago) |
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Nice Tek I'm try this tomorrow, I am just waiting for my silicone to dry. Could I make a jar of this up and keep it in the fridge and use it as and when needed? would I need to cover the filter or anything?
Also I have made a few of these lids up I'm guessing they will be fine for my grain jars? Starting first WBS grow this weekend
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OregonChronic
Iron, Lion, Cyans
Registered: 09/12/10
Posts: 1,970
Loc: Scoping Woodchips
Last seen: 11 years, 2 months
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Re: Multiply Spores Tek for n00bs [Re: hawkington]
#15454247 - 12/02/11 11:39 AM (12 years, 4 months ago) |
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Yes the lids can be used for normal grain jars too. And yes you should wrap the top in alum foil and store the spore water in a fridge. You can wrap the whole jar if you wanna. This is how I prep all my lids.
I've had problems when I make the tyvek square too small. It shrinks and may cause problems with the micropore seal. But I prep many jars like this with no problems. And just a few days ago RR informed us that postal tyvek such as in this tek, will shrink or enlarge when PC'd, making it either shrink and give less GE or it can expand and let spores in.
RR says to use either synthetic filter discs or paint overalls tyvek as its much thicker and won't distort when PC'd. But I've used this exact tek with postal tyvek with no problems on ALL my grain jars. Just make sure the silicone fully cures before you perform the tek. If it makes the jar smell like vinegar when put on then let cure longer.
-------------------- Wavy Cap Cyan Spore Prints for Trade/Sale
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SomeGuy
I feel better now :)
Registered: 04/18/10
Posts: 7,496
Loc:
Last seen: 9 years, 10 months
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this is how I do my grain jars, except I use clear tape instead of micropore, because the tyvek is the filter and the micropore isn't tight enough to stop bacteria/spores. I have found I have more success with 2 layers of postal tyvek, I just put one on, tape it, and then the next one and tape it
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OregonChronic
Iron, Lion, Cyans
Registered: 09/12/10
Posts: 1,970
Loc: Scoping Woodchips
Last seen: 11 years, 2 months
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Re: Multiply Spores Tek for n00bs [Re: SomeGuy]
#15454333 - 12/02/11 12:08 PM (12 years, 4 months ago) |
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Never thought of that. It would be way better to use clear tape as long as it doesn't melt in the PC. Might edit the tek to use two layers of micropore for extra safety. Though I have had no contams yet using only one layer of micropore, I can see how something might squeeze in its not so tight pores.
-------------------- Wavy Cap Cyan Spore Prints for Trade/Sale
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SomeGuy
I feel better now :)
Registered: 04/18/10
Posts: 7,496
Loc:
Last seen: 9 years, 10 months
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something I found out.....that "frosted" tape, transleucent, shrinks really bad and pulls up, so only use clear, "transparent" tape it holds a lot better
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hawkington
Registered: 11/23/11
Posts: 29
Last seen: 12 years, 3 months
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Re: Multiply Spores Tek for n00bs [Re: SomeGuy]
#15454512 - 12/02/11 01:07 PM (12 years, 4 months ago) |
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wicked. cheers for the advice.
I did actually use Tyvek that I cut out of my overalls, just the one layer. Then I stuck it down with 2 layers of micro pore tape. My tape isn't 3M brand and doesn't feel that sticky Im dubious about it holding up in the PC but we will see. If the tape fails I will just stick the Tyvek down with silicon.
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sillycube
hatter
Registered: 10/19/11
Posts: 54
Loc: gone to spore
Last seen: 9 years, 5 months
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Re: Multiply Spores Tek for n00bs [Re: hawkington]
#15454555 - 12/02/11 01:20 PM (12 years, 4 months ago) |
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seems smarter to go ahead and use those 2-5 ccs of spores to start a couple jars of actual liquid culture, since its easier than even PF tek and LC colonizes like twice as fast (as multispore)
also i like to just put tyvec over the whole damn lid and let the rim hold it on, andjust smush it into the silicone for the inoc port.
-------------------- "when i was younger, i heard about the decline of western civilization, and i decided it was something i would like to make a contribution to" - George Carlin.
Edited by sillycube (12/02/11 01:26 PM)
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hawkington
Registered: 11/23/11
Posts: 29
Last seen: 12 years, 3 months
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Re: Multiply Spores Tek for n00bs [Re: sillycube]
#15454602 - 12/02/11 01:32 PM (12 years, 4 months ago) |
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Yeah I'm going to get a LC going as well. Just need to make the best use of what I have left as a smashed pretty much a whole 10ml syringe on my first 10 PF cakes which seems like a bit of a waste of £15 now when it could have gone alot further. I guess i just wanna do as much experimenting as I can at the moment and see what works out for me. SO MUCH TO DOOOO
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OregonChronic
Iron, Lion, Cyans
Registered: 09/12/10
Posts: 1,970
Loc: Scoping Woodchips
Last seen: 11 years, 2 months
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Re: Multiply Spores Tek for n00bs [Re: sillycube]
#15454678 - 12/02/11 01:54 PM (12 years, 4 months ago) |
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It's supposed to be designed for noobs who don't know much about LC yet. I can throw in a link of Agar's glc tek in the OP too just to show how easy that is.
I was also gonna create a tek of mixing a little bit of brf and simply put a bit of it in the bottom of a jar like Agars tek but with brf. And use a filtered lid with silicone port so no dry verm is needed.
Just inoc it with MS let colonize and aspirate sterile water into it just like glc tek, but with brf. So no PC is needed just steam sterilize everything for 90mins. Wrap syringes in alum foil and steam sterilize as well.
I'm gonna try that out and of course it will work. Just because it would be real helpful to noobs without PC yet.
-------------------- Wavy Cap Cyan Spore Prints for Trade/Sale
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blackout
Registered: 07/16/00
Posts: 5,266
Last seen: 5 months, 10 days
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I have diluted spores sort of similar to this, but did not bother with a filter and only used 1 syringe instead of buying loads of normal or empty ones. Or just used a 60ml syringe as my extra.
If you have loads of jars I think you can request dark syringes from some vendors (not sure which ones these days).
So I would get distilled water (break some built up frost off your freezer) and sterilize it with a jar with a silicone injection port. I would sterilize this in a microwave or PC. I would close it up while still hot. If doing it in a microwave I would have the cap on loose (don't bother showing your ignorance by lecturing me about metal caps in microwaves, educate yourself). I would put them in a plastic bag loosely tied, NO TINFOIL. I would leave them about 5mins after they were finished boiling, I put them on low for about 30mins after they have started to boil.
While still hot I close the cap and shake it, to get any airbourne contams into the water and washed off the side walls of the jar if they landed there. Then wrap in a towel or blanket and allow to cool, the towel insulates it so keeps it warm which I believe will knock out most contams if any did get in.
This creates a vacuum in the jar -no problem. Get your syringe and inject it (after flaming & wiping). Now the entire syringe or almost the whole thing gets drawn into the jar automatically because of the vacuum. This usually depends on the head space you leave in the jar.
Now take the syringe out, hold a lighter to the needle at the end and slowly draw air in. Any contams in the air trying to pass through the needle should be killed. Now stick it in the jar, if the vacuum sucks all this air in again you might have to repeat this until the vacuum is gone.
Now you can suck up some spores and keep repeating the "flamed air" injections, wiping the port as you go, the needle is already hot from the air sucking procedure.
I would highly recommend getting 60ml or larger syringes, as this really speeds things up and the less injections you need the less risk of contams in the process.
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