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How to Make Liquid Culture | Liquid Culture Lids | Take a Live Culture | Inoculate a Spawn Bag - YouTube
Channel: Boomer Shroomer
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hi ashley here and today i'm going to聽
show you how to make liquid culture聽聽
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liquid culture is a great way to speed聽
up colonization and preserve genetics聽聽
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one downside to lcs is you can't see contamination聽
so checking viability on agar is best聽聽
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to start we need to make the lids these lids聽
are made of pp5 plastic which means that you聽聽
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can sterilize them in a pressure cooker they聽
are 90 millimeters in size and fit on a 32聽聽
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ounce mason jar these plastic lids are great聽
because they don't rust like the metal ones do聽聽
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now we need to drill two holes one for聽
the red injection port and one for the聽聽
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gas exchange filter for the gas to exchange聽
port i'm using a 5 and 30 seconds drill bit聽聽
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and we just drilled them directly on聽
the mason jar and it worked really well聽聽
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then insert the 13 millimeter filter聽
into the hole and it should be a nice聽聽
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snug fit then for the self-healing injection聽
port i used a half inch forstner drill bit聽聽
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then screw the lid on a mason jar and drill the聽
larger hole for the self-healing injection port
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finally let's insert the聽
self-healing injection port聽聽
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just pinch it and try to push it through the hole聽
and then from the other side you can just kind of聽聽
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pull with your finger or your thumb around聽
the edges and then it'll just pop into place聽聽
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now that our lids are finished we聽
can start making our liquid culture聽聽
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first weigh out eight grams of your liquid culture聽
media i am using a pre-mix from shroom supply but聽聽
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any extra light malt extract will work next we聽
need to put our magnetic mixer in the mason jar聽聽
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it's very important that you put the mixer聽
in before you sterilize your liquid culture
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then add your liquid culture media to the mason聽
jar i'm using a 32 ounce wide mouth mason jar聽聽
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then add 600 milliliters of water to the mason jar聽聽
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then thoroughly mix on a mixing聽
plate until it's completely dissolved
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next we need to strain the聽
mix twice with a coffee filter聽聽
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this helps remove a lot of the extra sediment聽
sitting at the bottom if you don't get all聽聽
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of the extra sediment at the bottom don't聽
worry the mycelium will eventually eat it聽聽
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i like to filter the liquid culture twice because聽
this gets about 80 percent of the sediment at the聽聽
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bottom and it comes out very clean and clear which聽
makes it a lot easier to see the mycelium growth聽聽
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then put the lid on and give it one last聽
mix and now we're ready to sterilize it
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next place a piece of aluminum foil over the lid聽
now we are ready to sterilize our liquid culture聽聽
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next let's prepare the pressure cooker place聽
the cleaning rack at the bottom for a spacer聽聽
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and then add around 8 to 10 cups of water聽
i tried to cover the spacer at the bottom聽聽
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then put your liquid culture mix in聽
the pressure cooker and close the lid
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then sterilize for 45 minutes at 15 psi after聽
45 minutes turn off the stove and release the聽聽
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pressure finally remove the liquid culture from聽
the pressure cooker and let it cool down to room聽聽
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temperature cooling it down to room temperature聽
can take anywhere from 12 to 24 hours if you add聽聽
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a live culture or spores when the liquid culture聽
is too hot you can end up killing the mycelium聽聽
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after it's cooled to room temperature it's聽
ready to go and now we can add our live culture聽聽
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now it's time to add our live culture it's聽
best to be in front of a laminar flow hood聽聽
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or in a sterile environment first you want to聽
pick a freshly harvested mushroom i try to pick聽聽
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the biggest strongest looking mushroom from聽
the flesh in order to get the best genetics聽聽
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first carefully open the syringe and the needle聽
and then carefully screw on the needle to the聽聽
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syringe trying to face everything towards聽
the laminar flow being as sterile as possible
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next you want to insert the needle into the red聽
injection port and then suck up some of the liquid聽聽
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culture you're sucking up some of the liquid or聽
culture because when you take the live culture聽聽
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as a biopsy in the needle you need something聽
to basically push it through the needle next聽聽
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we're going to take our life culture so聽
you're going to want to take your mushroom聽聽
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and then carefully split聽
the mushroom down the middle
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and let it sit towards the laminar flow just聽
in case you can't get a biopsy from one you聽聽
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can use the other then remove the needle from the聽
mason jar with the liquid culture in it and then聽聽
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take a biopsy of your liquid of your mushroom聽
and i try to use the bottom of the mushroom聽聽
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um i heard that it could be the strongest part of聽
that mushroom then you kind of want to twist your聽聽
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needle a little bit until you can get a chunk聽
in there took me a really long time i struggled聽聽
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then once you get a chunk you'll insert聽
the needle back into the red injection port聽聽
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and then push the live culture through the needle聽
into the mason jar okay you can see a small piece聽聽
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of the live culture right there you just need a聽
really small piece you don't need anything big聽聽
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there's also another piece i actually got two so聽
that's to be sufficient then over the next couple聽聽
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of days you will start to see your mycelium聽
grow here is the next day all of that at the聽聽
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bottom is actually the sediment from the liquid聽
culture media and it looks like contamination聽聽
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but it's actually not and over the next couple聽
of days the mycelium actually starts to eat it聽聽
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so it's not a big deal you'll want to mix up your聽
liquid culture every day just kind of hand mix it聽聽
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and then we can start to see a chunk it'll聽
start to float around here uh it takes me a聽聽
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minute okay here is a chunk of the mycelium and聽
you can see a tiny bit of growth on the outside聽聽
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of it that's the first day of growth it looks聽
pretty good it's not too bad ah that's a better聽聽
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angle that looks really good right聽
there you can see it all the way around
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then this is the fifth day growth and each day聽
like i just want to give it a quick mix and you聽聽
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can see that the sediment is slowly getting聽
eaten by the mycelium and this stuff grows聽聽
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really fast so it's already starting to break up聽
into you know five to ten pieces and definitely聽聽
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noticing growth on all those pieces so聽
everything that you see there is mycelium growth
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okay this is day six and the mycelium is聽
really starting to clump there together
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and this is day seven and i think that we have聽
enough to take a test to check for contamination聽聽
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so that's the next thing that we're going to聽
do is go ahead and test to see if our liquid聽聽
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culture is clean and i'm going to test on agar聽
now we're ready to test our liquid culture on agar聽聽
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first clean off the red self healing聽
injection port with an alcohol pad聽聽
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then start to prepare your syringe open the聽
syringe and the needle facing the laminar flow聽聽
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trying to be as sterile as possible and then聽
screw the top of the needle onto the syringe聽聽
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next insert the needle into the聽
red self healing injection port
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then carefully try to suck up some of the liquid聽
live culture in the mason jar and this is actually聽聽
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a lot harder said than done i try to swirl the聽
live culture up to the top of the mason jar聽聽
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which makes it a little bit easier聽
to try to get but it takes some time聽聽
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working the mycelium around in the聽
mason jar until you actually get聽聽
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a good chunk inside of the syringe聽
in order to test it on the agar
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after you get a decent chunk in the syringe next聽
test it on the agar so here you can see that聽聽
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there's quite a bit of liquid in the syringe and聽
i don't really think that you need all that liquid聽聽
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so i actually try to squirt some of it out and聽
only get the piece of culture on the agar if it's聽聽
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too liquidy that can actually cause additional聽
contamination so you can see me here like i really聽聽
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get most of the liquid until the live culture is聽
right at the tip and then squeeze it onto the agar聽聽
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but it's okay if you test you know the the聽
actual water in the syringe too because we聽聽
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want to make sure that that's contam free as聽
well then with these all live cultures the聽聽
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next day you should see mycelial growth it's聽
super fast these live cultures are so quick聽聽
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so there you go go ahead and wrap up your petri聽
dishes with parafilm and you guys are ready to go
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okay this is the next day for my agar trace sorry聽
that the video quality is so dark um but you can聽聽
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see that the mycelium has already started to grow聽
and has a pretty good chunk started um and no聽聽
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contamination and then this is five days later and聽
the mycelial growth is definitely a lot bigger and聽聽
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still no contamination so we are good to go yay聽
after checking for contamination now we are ready聽聽
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to inoculate our grain spawn bag with our liquid聽
culture first i recommend being in front of a聽聽
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laminar flow hood or in a sterile environment then聽
sanitize everything before you start this liquid聽聽
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culture has been colonizing for about three weeks聽
and it's pretty much covering the whole mason jar聽聽
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which is the perfect time to go ahead and take聽
a syringe of it and inoculate our grain spawn
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next we need to prepare the syringe first聽
open the syringe and then the needle聽聽
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facing the laminar flow and then聽
screw the needle on top of the syringe
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then you will want to insert the needle聽
into the self-healing injection port
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and suck up some of the liquid culture聽
into the syringe filling the syringe
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then carefully remove the needle聽
from the self-healing injection聽聽
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port and you should see a bunch of liquid聽
culture in there you can see it in there
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and carefully inoculate the grain spawn bag聽聽
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you'll want to put about two to four cc's聽
of liquid culture in your bag kind of聽聽
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inserting it in all the corners of the bag careful聽
not to puncture through the back of the bag
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then give the bag a quick聽
mix and you are ready to go聽聽
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lastly you can take a piece of micro pore tape and聽
just cover the self-healing the black self-healing聽聽
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injection port on the spawn bag then the next聽
day you will see growth so this is day one聽聽
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the next day after i inoculated and there's just a聽
tiny bit of mycelium growth but definitely growth
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and then this is day two
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and this is day four and you can really see聽
that the bag is really starting to take to聽聽
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the liquid culture and then there are spots聽
of mycelial growth popping up all over the bag
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and this is day six so over the next聽
30 days you're going to see your bag聽聽
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colonized and then once it fully colonized聽
you can move it into your mono tub聽聽
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thank you guys so much for watching and if聽
you have any questions please feel free to聽聽
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put them in the comments you can reach out to me聽
on instagram or facebook also please subscribe to聽聽
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my new channel ashley schumer where i'll be聽
posting new mushroom content soon thank you
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