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A *beginner friendly* method for making Liquid Culture * 56
    #5238137 - 01/29/06 10:52 AM (18 years, 2 days 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 02:14 AM)


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Re: A foolproof method for making Liquid Culture [Re: TheDemon] * 2
    #5238206 - 01/29/06 11:22 AM (18 years, 2 days 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 08:28 PM)


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Re: A foolproof method for making Liquid culture [Re: blackout] * 1
    #5238239 - 01/29/06 11:33 AM (18 years, 2 days 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: Dribblesmeer] * 3
    #5239856 - 01/29/06 07:50 PM (18 years, 2 days 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: kevinmatk] * 1
    #5240012 - 01/29/06 08:30 PM (18 years, 2 days 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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Re: A *beginner friendly* method for making Liquid Culture [Re: weedpreference] * 1
    #5240311 - 01/29/06 09:37 PM (18 years, 2 days 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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Re: A foolproof method for making Liquid culture [Re: BlistCaster] * 2
    #5243233 - 01/30/06 06:02 PM (18 years, 1 day ago)

Quote:

BlistCaster said:
Thanks for the tutorial! I was just wondering why someone can't just add a lot more honey to allow for more nutrients in the water and thus more mycelium...?




More is not better mate - honey is a natural preservative, and if you make the concentration too strong it will inhibit the growth of the mycelium.

Quote:

HyphalTryptamin said:
Maybe your honey is weak - its only usually a tablespoon to 250 ml when we're talking about powdered ingredients - like dry light malt extract and dextrose monohydrate. For syrup-like substances like proper Barley malt extract and honey - 1 teaspoon is plenty man :thumbup: - good work though nonetheless :laugh:




I do agree a teaspoon is enough - but I've always had much better results adding just a *little* bit more - it makes the mycelium nice and thick.... and I think honey is a lot more forgiving than than other syrups.

Maybe if your experimenting with it for the first time try a teaspoon and work up from there.

I love the fact honey has so many fantastic natural healing qualities... its like the 'healthy' option for your LC - and its also great on toast in the morning!

Quote:

psychedelix said:
I'm going to try this.  A while back when I actually had the heart for this hobby, almost all my LCs failed, since I haven't attempted an LC stunt... but this looks like it's so easy that even if it fails, I won't be pulling my hair out stressing.




I wish you luck with your LC and I hope you 'revive' your passion for the hobby.

Quote:

psychedelix said:
Awesome tek... I say it deserves to get archived. :thumbup:




Thanx heaps mate! Maybe you could nominate it then?


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Re: A foolproof method for making Liquid culture [Re: License_to_Chill] * 1
    #5258161 - 02/03/06 02:20 AM (17 years, 11 months ago)

Get some water boiling on the stove for 15 mins to sterilise it, then slowly fill the needle with the still boiling water and shoot it back out into the sink (obviously not back into the pot!). Do this 5 or 6 times to clean out the inside of the syringe, then flame sterilise the tip on the flame on the stove - or wipe it with alcohol, or both.


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Re: A foolproof method for making Liquid culture [Re: License_to_Chill] * 1
    #5258177 - 02/03/06 02:35 AM (17 years, 11 months ago)

Thank you very much.... :wink:


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Re: A *beginner friendly* method for making Liquid Culture [Re: Blek] * 1
    #5258815 - 02/03/06 11:06 AM (17 years, 11 months ago)

Quote:

Picking4Peace said:
In other words, should I worry about there being contaminants on the mycelium, or is it ok to just break off a chunk and put directly into the honey/water jar?




Yes you can use a peice of myceiulm from a cake, but be carefull of contams. Cut a peice out of the centre of the cake with a sterile knife. Alternatively, cut open the inside of the stem and use a small chunk of the flesh, to 'clone' a mushroom.


Edited by _OttO_ (02/03/06 06:27 PM)


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Re: A foolproof method for making Liquid culture [Re: Oatman2000] * 1
    #5260264 - 02/03/06 06:34 PM (17 years, 11 months ago)

Quote:

oatman2000 said:
any differences between honey and karo... in your opinions?




Karo LIGHT makes a clearer end product which makes it a lot easier to see the mycelium (and any contams). I just use honey because I always have some on hand in the house.


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Re: A foolproof method for making Liquid culture [Re: CaptainKirk] * 1
    #5260360 - 02/03/06 07:05 PM (17 years, 11 months ago)

That looks really healthy mate. I love the fact you dont need to hassle about with making spore syringes and makes it easy to keep growing from your own prints.


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Re: A *beginner friendly* method for making Liquid Culture [Re: sever] * 1
    #5268314 - 02/06/06 01:14 AM (17 years, 11 months ago)

Probably best to start a new thread for something like this, and without being able to say exactly what went wrong - I can say that you needed to sterilise the jar again after the honey was added.

The smell *might* have been a sign of bacterial contamination.


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Re: A *beginner friendly* method for making Liquid Culture [Re: FractalXplora] * 1
    #5344592 - 02/27/06 09:22 AM (17 years, 10 months ago)

Good luck and make sure you post your results!


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Re: A *beginner friendly* method for making Liquid Culture [Re: Aloysius] * 1
    #5454136 - 03/29/06 07:56 AM (17 years, 9 months ago)

Quote:

Aloysius said:
This might be worlds stupidest question, but if I took some tissue from the inside of a shrooms' stem, and tossed that in a pot of water/honey solution (everything as sterile as possible of course)..

Would I see some growth?




Yep, sure would! You would be 'cloning' the mushroom the peice of stem came from. :thumbup:

Quote:

Bamaman said:
Question:

I saw it said that syringes of your lc will keep in the fridge for months. What about leaving it in the jar, baby bottle, etc...? I would assume that it would "keep" in the original LC jar as well, right?

It would be alot less conspicuous for long term storage in your fridge.




Definately - I dont remember saying to suck it up and store, I just keep the jars of mycelium in the fridge - and suck some up as you need to use it.

This is still working fine for me and still hasnt contammed yet. I have been having casing contams lately - I have been a bit lasy and too busy - but its nothing to do with the LC and I still have never had one jar contaminate.

I always open air inoculate the LC by just scraping up some spores with a knife tip and dropping them into the jar.


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Re: A foolproof method for making Liquid culture [Re: kilroy] * 1
    #5698675 - 06/01/06 07:37 AM (17 years, 7 months ago)

Well I have never had one contam on me yet. Not sure if I am just careful with the process or if its not as susceptible to contamination as ppl say, but yeah - never a problem with many open air LC innoculations from a spore print.

I had a couple of LC's at the back of the fridge freeze solid on me the other day (the fridge was too low). I defrosted them and they were still viable - eight jars all colonised without a hitch.


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Re: A foolproof method for making Liquid culture [Re: kilroy] * 1
    #5698793 - 06/01/06 08:46 AM (17 years, 7 months ago)

Use a larger amount of LC to inoculate your jars, and you reduce the time that any potential contams can take hold. I use about 7cc of LC per jar.


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Re: A foolproof method for making Liquid culture [Re: tallgreen] * 1
    #5761847 - 06/17/06 09:22 PM (17 years, 7 months ago)

Quote:

tallgreen said:
I just did this tek 2 days ago and now there is lots of stuff (mycelium I hope) settled at the bottom. It clouds the jar when shaken. So I think it's good to go. My first LC. Thanks _oTTo_!!





The stuff that has settled at the bottom is sediments from the honey - not mycelium, but keep swirling it every day and it dissolves again. While the mycelium grows and consumes the nutrients, the sediment seems to dissapear.

Quote:

hippie_cune said:
how long can one of these last?




In the fridge, many months.


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Re: A foolproof method for making Liquid culture [Re: acidbuba] * 1
    #5802365 - 06/29/06 07:55 AM (17 years, 7 months ago)

IMO - I think that you are trying to be too technical with it - for your first attempt, forget the spot of silicon over your hole.

Forget about the paperclip and the PC - just steam sterilise your LC.

Just cover the hole with the foil, and dont forget to shake well after making and sterilsing your LC - this aerates your mixture and encourages growth.

Then Innoc with a scraping of a print or a bit of a spore syringe, and re-cover the hole with the foil lid.


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Re: A foolproof method for making Liquid culture [Re: Blutjager] * 1
    #5805929 - 06/30/06 03:48 AM (17 years, 6 months ago)

Yeah, good advice Blutjager. You should never be stingy with your LC - just use the remaining bit to make more, or, just take a print or two from your harvest and use that. Even better, take a chunk of tissue from your best looking shroom and use that to start a new LC - then it will be cloned.

Remeber, more LC = faster colonisation.

Inject 10cc and see how you go!


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