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Offline_OttO_
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A *beginner friendly* method for making Liquid Culture * 45
    #5238137 - 01/29/06 12:52 PM (14 years, 4 months ago)

Hey everyone - I'm writing this because I see a lot of questions being asked about Liquid Culture. My first ever grow started off with a LC and I am convinced it should be *Edit to remove 'all or nothing' statement something everyone should try at least once, even people who are still using the PF tek who don't have access to a Pressure cooker.

You don't need expensive equipment and generally most people will have access to these materials already around the home, which should help even the most nervous beginner to get off their bum and have a crack at playing with mycology!

*Edit to include motivational speech* NB: This method is not technically completely sterile, but if you have a clean Spore print/spore syringe, and follow the steps exactly, you should have no problem. If however you do encounter contamination problems don't be put off - try again! Even professionals still make mistakes and you can only get better with experience!! :smile: :smile: :smile: The same goes for anything you do in life... :wink:

A photo says a thousand words, and there doesn't seem to be any simple teks around that are well documented with pictures describing each step of the process for the absolute beginner .

What I do is no way new or fantastic, I simply use a mixture of a few different teks found on this site, and I don't seek praise for following other people's ideas. I just want to demonstrate to the new cultivator how easy this is, and outline a step by step process that works for me for them to follow, in the hope more people try this step in their cultivation practice.

Materials:

A small, clean glass jar with a metal lid - I use an empty salsa jar -



A pot with a lid - *Edit* Or a Pressure cooker, if you have one.



Some clean tongs -



Some foil -



Some disinfectant spray with a high alcohol content and some paper towel -



And some honey (the clearer the better - not 'cloudy' honey) *Edit to include a a clearer option* Pure maple syrup, or a combination of honey and maple works really well for me to get a clearer final product -



Some rubber latex gloves, and lastly;

A clean spore print or spore syringe.

Method:

Punch a hole in the lid of the jar with a nail with a hammer and nail -



Fill the pot with cold water and submerse the jar and lid in it -



Bring this to the boil (never drop the cold jar straight into boiling water or it will crack). The purpose of this is to partially sterilise the water and the jar -



Set a timer for 30 minutes, crack a beer and go and check www.shroomery.org

After 30 minutes, clean the tongs and the bench top you are going to use with the alcohol spray and paper towel. Use the tongs to lift the jar out of the boiling water and place it on your bench -



making sure you keep the jar 3/4 full of the boiled water. Be careful - its hot! - 



Add a tablespoon of honey to this. Work on using a tablespoon of honey for every 250 mls of water you have - *Edit to reduce the amount of honey added and introduce a new option for a clearer LC*  I have since found that about a teaspoon per 200 mls of water is a better ratio to start with, and for an even clearer final product, substitute half or more of the honey for pure maple syrup to reduce sediment related issues from some brands of honey. Half honey and half maple is my favourite so far!



Use the tongs to take the lid out of the boiling water, then using a tea towel (hot!) put the lid on the jar. Leave the pot of water still boiling on the stove -



Tear off some foil and crumple it tightly over the lid of the jar. At this point in time I like to put a 'dot' on the foil with a permanent marker pen above the hole that is in the lid, so I can remember where the hole is -



Place this jar back in the water thats still boiling on the stove - you want the water to come about half way up the jar - if need be, just tip some out of the pot -



Place on the tight fitting lid -



Reduce the heat to a low simmer, set your timer for another 30 minutes then crack a beer and go and check www.shroomery.org

When the timer goes off, turn off the stove and DON'T PEAK - just leave it with the lid on the pot to cool overnight. Inside the pot is now a sterile environment and opening the lid is an invitation for contaminates.

*Edit to include pressure cooker advice* Alternatively, you can use a pressure cooker at 15psi for 15-20 minutes, if you have one. This is the preferred method!

Inoculation:

With a spore syringe -

The next day when its cool, simply remove it from the pot to a pre-cleaned bench top and have your spore syringe ready -



*Edit to change amount of spore solution, but if your syringe is clean, the more you inject, simply the the quicker it will grow* Flame sterilise the tip of the needle then peel back the foil on the jar to expose the small hole you made in the lid. Insert the syringe and inject 1-2CC of spore solution -



Replace the foil.

With a spore print -

Ok - this is a little less sterile but still my preferred method because it helps you be more self sufficient by allowing you to start the culture directly from your own prints and cuts out the step of making a spore syringe (a step which only adds another chance for contams to enter anway!) - I always use this method and I have not had one contamination problem yet.

*Edit to clarify the importance of still air* Close all windows and doors a few hours before you start working. This is really important as you want as little air movement as possible where your working as this stirs up contamination spores. You also want to make all you; movements swift but smooth, in order to minimise the air movement/stirring up contamination spores.

*Edit to include sanitised gloves* Now put on some rubber gloves and sanitise them with some a squirt and rub together of your alcohol spray. Then clean and sanitise your bench top as best you can your with the alcohol spray and paper towel.

Get ready your LC, your spore print and a sharp knife on your bench top -



Flame sterilise the tip of the knife -



While this cools, crack the lid of the jar slightly - but dont open it completely , and get your spore print ready.

Once the knife is completely cool, use it to scrape up some spores. Holding your breath, in one smooth motion, scrape up as many spores on the tip of the knife as you can.



Lift the lid of the jar open *just a crack* and insert the tip of the knife, dropping the spores off into the solution - a little tap on the glass will help them fall off, and you should see some land on the top of the liquid -



Replace the lid.

After inoculation:

Place your finger on the 'dot' over where the hole in the lid is and swirl gently. This is to distribute the spores in the solution and to encourage growth.

Label with the date and strain etc. and incubate this in a dark place at 29C or 82F, giving it a gentle swirl once a day to encourage growth.

After a few days you will notice some small strands of mycelium forming, and it will be ready completely after about a week. You will notice the liquid will become much lighter as the mycelium uses up the nutrients in the water, and will come to a point where it will stop growing -



To use simply agitate it slightly to loosen up the mycelium, peel back the foil and insert a sterile syringe to suck up as much mycelium as you can - use this as normal to inoculate any substrate.

One can inject larger amounts of this than you would normally (like 2cc at each injection site) for dramatically sped up colonisation times.

I firmly believe once you try this method you will never inoculate with spores again!

If there is anything anyone thinks I need to add feel free to comment. Thanx and good luck!


Edited by _OttO_ (04/14/14 04:14 AM)


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InvisibleTheDemon
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Re: A foolproof method for making Liquid culture [Re: _OttO_] * 1
    #5238145 - 01/29/06 12:54 PM (14 years, 4 months ago)

FDSFGS

Nice tutorial!


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OfflineNunbuh_Chrubble
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Re: A foolproof method for making Liquid culture [Re: TheDemon]
    #5238180 - 01/29/06 01:11 PM (14 years, 4 months ago)

Very nice! It's good that you've proven this can be done without a PC! It makes the whole idea much more accessable to timid newbs.

I would say though, that you made your solution quite dark with honey! I literally put as small of a drop as possible in about a pint of water. It stays crystal clear, making it easier to tell if the myc is white or some other unwanted color, and the amount of growth is quite sufficient considering that most of the LC will probably expire before I can use it all!

I guess some just like it nice and thick... but I think it should be emphasized that it's almost impossible to put too little syrup in there, but it's quite possible to put too much. Newbs get all stressed about measuring out the ratios for their LC when it is completely unneccessary IME. Perhaps if you were doing quart or gallon sized LC's then you would probably want to measure it out, but for the amateur noob guestimating should be fine.


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Offline_OttO_
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Re: A foolproof method for making Liquid Culture [Re: TheDemon] * 1
    #5238206 - 01/29/06 01:22 PM (14 years, 4 months ago)

Yeah I never measure it either but thought I should put a specific measurement for beginners to follow - you get good at eye-balling the right amount after a while, and this is roughly how much I use.

I do like a thicker culture though, so I tend to go a bit heavier on the honey - like I said however, the solution goes almost completely clear after a week when the nutrients are used up.

Quote:

Nunbuh_Chrubble said:
and the amount of growth is quite sufficient considering that most of the LC will probably expire before I can use it all!




Use more LC!! Ive had phenominal results by injecting larger amounts of LC into Bird seed. Dont let it go to waste, make your substrate a little drier and inject 10 or 15cc of LC per jar and watch how fast it goes!


Edited by _OttO_ (01/29/06 10:28 PM)


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Offlineblackout
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Re: A foolproof method for making Liquid culture [Re: _OttO_] * 1
    #5238227 - 01/29/06 01:28 PM (14 years, 4 months ago)

Nice. I put 2 holes in my jar and cover with silicone sealant on both sides. this allows you to shake the jars really well to aerate and break up the myc. The second hole lets me put a filtered syringe in so it equalises the pressure when sucking out big amounts of liquid.


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Offline_OttO_
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Re: A foolproof method for making Liquid culture [Re: blackout]
    #5238239 - 01/29/06 01:33 PM (14 years, 4 months ago)

An excellent addition Blackout - only in this example Im trying to keep it as simple as possible for the noob cultivator. You never get a build up of pressure either as there is room around the syringe tip for air to enter the jar - I know this isnt going to be sterile but I have never had a contam regardless.

BUT - once they have the hang of it - silicone over the holes and filter syringes are highly recommended to make your LC easier to manage.


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Re: A foolproof method for making Liquid culture [Re: _OttO_]
    #5238278 - 01/29/06 01:47 PM (14 years, 4 months ago)

Yes simple is best, everybody can get a jar! the hole is so small there wont be much room for airborne contams. A lot of my LCs are failing lately, I discovered a large patch of mould growing on a wall in my room due to condensation, I have since bleached down the walls and sparyed lots of disinfectant everywhere. I hope you good luck in avoiding contams continues.


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Offlinescatmanrav
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Re: A foolproof method for making Liquid culture [Re: blackout]
    #5238299 - 01/29/06 01:56 PM (14 years, 4 months ago)

You can also get anti fungal paint, if the problem persists..

Nice simple tek, well setup. Pretty damn easy too.


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Anyone looking to start bulk tubs/mono tubs/shotgun hybrids? Good tubs to use..
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Re: A foolproof method for making Liquid culture [Re: scatmanrav]
    #5238359 - 01/29/06 02:17 PM (14 years, 4 months ago)

I've heard throwing in a couple of marbles helps break it up better when u shake your solution. Do you consider this helpful?


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OfflineHengman
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Re: A foolproof method for making Liquid culture [Re: Dribblesmeer]
    #5238384 - 01/29/06 02:26 PM (14 years, 4 months ago)

AWesome!!! i also agree that liquid cultures are essential. it can make LOTS of syringes with mycelium solution, rather than spores so you can skip the germination stage.


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OfflineCubenisseur
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Re: A foolproof method for making Liquid culture [Re: Hengman]
    #5238610 - 01/29/06 03:22 PM (14 years, 4 months ago)

I am one of the newbs thats been looking for a good simple LC tek, and this is quite fitting.


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Re: A foolproof method for making Liquid culture [Re: Cubenisseur] * 1
    #5238946 - 01/29/06 05:05 PM (14 years, 4 months ago)

I was considering using some leftover spore solution to attempt a LC but I don't know how long it's good for...

Will it last until I'm ready to make more jars (maybe 1.5-2months tops)?


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Re: A foolproof method for making Liquid culture [Re: Newbie]
    #5238988 - 01/29/06 05:21 PM (14 years, 4 months ago)

Great post Otto!

I have several questions as I have been SOOO tired of MS inoculation:

1) What is a filter syringe

2) Once an LC syringe is created, how long will it last if kept dark and in a clean ziploc at room temp?
and
3) I use a PC so should I let the pressure go down then add the culture to the jar, then rePC for another 30 mins? I'd open it after the gauge went down but it will still be fucking hot.

Thanks guys


Edited by Atheist (01/29/06 05:22 PM)


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Re: A foolproof method for making Liquid culture [Re: Hengman]
    #5239029 - 01/29/06 05:33 PM (14 years, 4 months ago)

Beautifully executed, Otto, you are marking a trail for many to follow. Sharing info, methods, and teks is the best part of this forum, and you are constantly there. Thanks.

I've been doing almost exactly what you have illustrated here, until yesterday, when I decided to try the microwave, as in agar's baby bottle tek. One thing I can share now is to make sure that if you use a plastic jar that originally had a foil seal, make sure you scrape the last little bits of metal off the rim of the jar. I had a lightning storm go off in my microwave, followed by a tyvek fire. It was sorta pretty, but real scary. :eek: :crazy2:


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Re: A foolproof method for making Liquid culture [Re: Dribblesmeer] * 2
    #5239856 - 01/29/06 09:50 PM (14 years, 4 months ago)

Quote:

Dribblesmeer said:
I've heard throwing in a couple of marbles helps break it up better when u shake your solution. Do you consider this helpful?




It can be helpful, yes - but Ive never done it and I find just shaking the solution alone will break up the mycelium enough.

I think agitating it every day plays an important role - it stops the mycelium 'clumping' together and becoming hard to break apart.

Quote:

NewbieShroomie said:
I was considering using some leftover spore solution to attempt a LC but I don't know how long it's good for...

Will it last until I'm ready to make more jars (maybe 1.5-2months tops)?




Yes it will, just keep the syringe in a ziplock bag in the fridge. Remember it takes about a week for the LC to develop properly, so factor this time in. And if its ready before you want to use it - simply put it in the fridge. The LC will store for many months in the fridge.

You can also use a small piece of clean mycelium from a cake or a small, clean chunk cut from the inside of a fruitbody to start your LC as well.

Quote:

SpicyTunaRoll said:
I have several questions as I have been SOOO tired of MS inoculation:

1) What is a filter syringe

2) Once an LC syringe is created, how long will it last if kept dark and in a clean ziploc at room temp?
and
3) I use a PC so should I let the pressure go down then add the culture to the jar, then rePC for another 30 mins? I'd open it after the gauge went down but it will still be fucking hot.





1) A filter syringe is used when you have an airtight seal on your jar. When you insert your syringe to suck up some mycelium, a 'vacuum' is created making it hard to suck up the plunger. You need to allow some outside air to come in, but you want this air to be clean. A filter syringe is a syringe with the plunger taken out and the body stuffed with polyfill, and this is inserted in one side of the jar when you go to suck up mycelium - to allow clean, filtered air BACK into the jar to equalise the pressure.

This isnt a problem with my method as enough air can get in around the hole where you insert the needle, but its small enough that air-borne contams aren't a massive concern.

2) I only make and use them immediately, but you can keep your jar of LC in the fridge for many months and simply take it out and suck up some mycelium whenever you need to inoculate some jars.

3) If your using a PC then you can completely skip the initial boiling step of the process - a PC will kill everything and this extra step is only for steam sterilisers as an extra barrier.

To use your PC, simply fill the jar with boiling water from the kettle, add the honey and lid, cover with foil, and PC the whole thing for 30 mins then allow it to cool overnight. I have a PC as well, but I prefer to use the steam sterilisation technique as its just as easy and doesnt require me firing up the bulky cooker.

Good luck, and please post any results!


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Re: A *beginner friendly* method for making Liquid Culture [Re: _OttO_]
    #5239957 - 01/29/06 10:16 PM (14 years, 4 months ago)

Thank you so much! Very useful, I'll be using this method on my first grow in a few days.


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Re: A *beginner friendly* method for making Liquid Culture [Re: kevinmatk]
    #5240012 - 01/29/06 10:30 PM (14 years, 4 months ago)

Quote:

kevinmatk said:
Thank you so much! Very useful, I'll be using this method on my first grow in a few days.




Good luck! Please post back your results... :thumbup:


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OfflineTeRzMaStA
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Re: A *beginner friendly* method for making Liquid Culture [Re: _OttO_] * 1
    #5240214 - 01/29/06 11:03 PM (14 years, 4 months ago)

That's awsome. That should help out a lot of people. 5 shroom rating for you.


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Re: A *beginner friendly* method for making Liquid Culture [Re: TeRzMaStA]
    #5240248 - 01/29/06 11:16 PM (14 years, 4 months ago)

im trying this method right now. i was wondering if using a lot of spores in one jar would speed up growth.


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Offline_OttO_
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Re: A *beginner friendly* method for making Liquid Culture [Re: weedpreference]
    #5240311 - 01/29/06 11:37 PM (14 years, 4 months ago)

Quote:

weedpreference said:
im trying this method right now. i was wondering if using a lot of spores in one jar would speed up growth.




It sure would! Just be careful not to indroduce contaminates. If your using a spore print, scraping up about the equivilent size of a match head from the print works fine - but the more the merrier! Ive scraped up just over 1/4 of a print before and it speeds things up a fair bit.


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