How to Make Liquid Culture | Liquid Culture Lids | Take a Live Culture | Inoculate a Spawn Bag

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

Channel: Boomer Shroomer

Views: 403,127

Keywords: mushrooms, How to Grow Mushrooms, Growing mushrooms, Agar, liquid culture, how to make liquid culture, monotub, spawn bag, monotub tek

Id: hw0IFzXSV0c

Length: 13min 12sec (792 seconds)

Published: Thu Mar 25 2021