Let's start w/ making an LC lid...
You need a jar and lid that can be PC'ed, I used a vaccutainer injection port that I removed from a test tube but you can use RVT silicone, and I used a filter patch from a used bag (after it was thoroughly cleaned and disinfected w/ bleach) but a Whattman filter plug is ideal.
OK now you need to make a hole in the lid for the injection port. I start off using the smallest drill bit and gradually increase bit size till I get it to the desired size. There's two reasons for this, first the thinner bits tend to be sharper and can make a cleaner hole in the metal lid w/o tearing the metal and ruining your lid like the duller thicker bits would. Second, you don't want to accidentally make the hole too big. I try to make it just big enough so I can barely fit the port in, making it snug and as tight a fit as possible.
Next, you need to make a hole for FAE...
You may also opt not to make an FAE hole...
Now you need to cut the filter patch to your desired size. I cut mine in half, then took one of the halves and cut again in half giving me 2 squares which is what I'll be using (2 layers, one square on top of the other)...
It's time to get your silicone ready...
Put some of the caulk around the hole that you made for FAE, giving it a wide birth so as not to accidentally fill the hole w/ silicone (that would defeat the whole purpose). Place the filter patch on, then put some more caulk lining the the perimeter of the square and place the second one on top. Take some excess caulk and put some more going from the edge of the patch to the lid. Also, put some caulk around the injection port where the port meets the lid (on both sides).
Let the silicone cure over night, the next day when you're ready to make the LC, clean it thoroughly before starting.
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So now you're ready to make the LC...
[My PC is busted so I'll be using a steamer]
Fill the pot w/ regular water and bring to a boil. While the water is heating up, make your measurements to figure out how much water you will need for your jar. Once the water is boiling turn off the heat and place the measuring cup w/ the DISTILLED water already measured out inside.
Use 1 teaspoon of brown rice syrup per 250ml of DISTILLED water.
Stir and mix it up. The reason I do this is b/c the heat from the warm water outside of the measuring cup helps to dissolve the syrup into the distilled water.
Fill up your jar and place it in the steamer. Bring the water to a boil again and let it sit for 90 min's.
Viola!!! Brown Rice Syrup LC!!! :stickdance:
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Inoculating the LC...
I inoculate my LC's w/ agar wedges, but you could do it w/ a spore syringe. OK so gather everything you're going to need/use and sterilize. [NOTE: These procedures should be done in front of a laminar flow hood or glove box.]
Loosen the lid, but don't open it yet. [If you're using a spore syringe, you don't need to do this, just plunge the needle through the port... after you sterilized it of course (needle AND jar/lid).]
Open up your plate of colonized agar and using your scalpel cut the desired sector. It doesn't have to be a huge piece... you can even put 2 small pieces in if you want...
Try to do this step as quickly as possible w/ the least amount of air movement. After you've put the wedge in, close the jar up tight and don't forget to label it...
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Don't worry about the cloudiness, it's just the brown rice syrup. After the myc starts to develop it will start to clear up b/c that's its food!!! lol This is what you'll end up with...
You need a jar and lid that can be PC'ed, I used a vaccutainer injection port that I removed from a test tube but you can use RVT silicone, and I used a filter patch from a used bag (after it was thoroughly cleaned and disinfected w/ bleach) but a Whattman filter plug is ideal.
OK now you need to make a hole in the lid for the injection port. I start off using the smallest drill bit and gradually increase bit size till I get it to the desired size. There's two reasons for this, first the thinner bits tend to be sharper and can make a cleaner hole in the metal lid w/o tearing the metal and ruining your lid like the duller thicker bits would. Second, you don't want to accidentally make the hole too big. I try to make it just big enough so I can barely fit the port in, making it snug and as tight a fit as possible.
Next, you need to make a hole for FAE...
You may also opt not to make an FAE hole...
Now you need to cut the filter patch to your desired size. I cut mine in half, then took one of the halves and cut again in half giving me 2 squares which is what I'll be using (2 layers, one square on top of the other)...
It's time to get your silicone ready...
Put some of the caulk around the hole that you made for FAE, giving it a wide birth so as not to accidentally fill the hole w/ silicone (that would defeat the whole purpose). Place the filter patch on, then put some more caulk lining the the perimeter of the square and place the second one on top. Take some excess caulk and put some more going from the edge of the patch to the lid. Also, put some caulk around the injection port where the port meets the lid (on both sides).
Let the silicone cure over night, the next day when you're ready to make the LC, clean it thoroughly before starting.
----------------------------------------------------------------------------------------
So now you're ready to make the LC...
[My PC is busted so I'll be using a steamer]
Fill the pot w/ regular water and bring to a boil. While the water is heating up, make your measurements to figure out how much water you will need for your jar. Once the water is boiling turn off the heat and place the measuring cup w/ the DISTILLED water already measured out inside.
Use 1 teaspoon of brown rice syrup per 250ml of DISTILLED water.
Stir and mix it up. The reason I do this is b/c the heat from the warm water outside of the measuring cup helps to dissolve the syrup into the distilled water.
Fill up your jar and place it in the steamer. Bring the water to a boil again and let it sit for 90 min's.
Viola!!! Brown Rice Syrup LC!!! :stickdance:
---------------------------------------------------------------------------------
Inoculating the LC...
I inoculate my LC's w/ agar wedges, but you could do it w/ a spore syringe. OK so gather everything you're going to need/use and sterilize. [NOTE: These procedures should be done in front of a laminar flow hood or glove box.]
Loosen the lid, but don't open it yet. [If you're using a spore syringe, you don't need to do this, just plunge the needle through the port... after you sterilized it of course (needle AND jar/lid).]
Open up your plate of colonized agar and using your scalpel cut the desired sector. It doesn't have to be a huge piece... you can even put 2 small pieces in if you want...
Try to do this step as quickly as possible w/ the least amount of air movement. After you've put the wedge in, close the jar up tight and don't forget to label it...
----------------------------------------------------------------------------------
Don't worry about the cloudiness, it's just the brown rice syrup. After the myc starts to develop it will start to clear up b/c that's its food!!! lol This is what you'll end up with...
