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