Bwpz
Well-Known Member
I like this idea, I could just keep a qt jar of the strain, and keep changing it out like every month-ish? I could just keep it in my incubator chillin? And still just 5cc's per jar, even a quart?
Bwpz' First PF Tek Attempt
- Thread starter Bwpz
- Start date
Bwpz
Well-Known Member
That's amazing information. I really appreciate you helpin me out there
I'm definitely going to take that route, I'll hit you up with a print when it comes time 
egon
Well-Known Member
if you already have mushrooms going you dont need to make an lc from spores, instead make it with some of the flesh from your biggest mushroom that is part of a cluster, that way you have a decent clone of that paticular mycelium, instead of random lc from multispore. and yeah lc colonize faster BUT they are not very good for brf cakes, better for wbs or popcorn, in which case its much faster.
Blackhash
Active Member
Who really knows if they colonize BRF slower than multispore?
So you can put a piece of mushroom matter into a LC and it colonizes it?
Check out this website for syringes.
http://www.mushbox.co/empty_BD_syringe.html
So you can put a piece of mushroom matter into a LC and it colonizes it?
Check out this website for syringes.
http://www.mushbox.co/empty_BD_syringe.html
egon
Well-Known Member
I know it does i've seen it with my own two eyes, i've even seen lc's fail to colonize brf, but they can colonize a few pints of popcorn or wbs in 12 days, seen that as well.
yeah there are ways you can make an lc from mushroom flesh its considered a clone and will preform much better than most multispore grows.
Bwpz
Well-Known Member
Do you have a tutorial on the one making the clone with the flesh? Or is it basically just throwing the flesh in the LC?
uromastyx
Well-Known Member
When I clone mushrooms I use a technique called tissue transfer using agar.
To do this you need petri dishes filled with agar this process can be as simple or as extravagant as you want it to be. There are tons of recipes to make your own agar, but I will type up a quick and easy one.
Materials:
1. Malt extract agar (MEA)
2. Water
3. Pressure cooker
4. Petri dishes
6. Canning jar able to hold 1.5-2L
7. Aluminum foil
8. Filter disc's or cotton wool
9. Glove box or flow hood
5. metal measuring spoon (needed if using peroxide)
6. 3% hydrogen peroxide (optional)
1. Add 50g of MEA into the canning jar, add 1L of water stir till its mostly dissolved, cover the jars opening with the filter disk and screw the outside metal ring back on the jar (if you don't want to use filter disc's just plug the opening with the cotton) now wrap the top with aluminum foil.
2. Place jar into pressure cooker (if you are planing on using peroxide toss the metal spoon in there also) add some water to the cooker and sterilize for 30min at 15psi.
Do not exeed 45min the MEA will start to caramelize and fungi doesn't grow to good on caramelized sugars.
3. Once you can open the pressure cooker again move the jar (and spoon) into your glove box or by your flow hood while still hot.
4. (When using peroxide: once the outside of the jar is manageable but still pretty warm around 120F-140F add 8ml of peroxide and swirl the jar to mix it, be careful not to over agitate the agar and create bubbles)
5. Open your sleeve (keep the sleeve we will need it again) of petri dishes and stack them inside your glove box or in front of the flow hood right side up (I stack mine in tens but it depends on how big your hands are and glove box)
6. Lift the first entire stack by the lid of the bottom dish and slowly pour just enough agar into the dish to cover the bottom. Repeat until all are filled. 1l per 20-30 100ml petri dishes.
7. Stack the completed dishes on top of each other and loosely cover with the sleeve they came in to ensure that they cool evenly this will also minimize condensation on the upper plates.
8. Allow plates to cool overnight
9. Now tape the sleeve shut and store them agar side up in a cool dark place.
now i need to go smoke and then il be back to write up the actual tissue transfer procedure
To do this you need petri dishes filled with agar this process can be as simple or as extravagant as you want it to be. There are tons of recipes to make your own agar, but I will type up a quick and easy one.
Materials:
1. Malt extract agar (MEA)
2. Water
3. Pressure cooker
4. Petri dishes
6. Canning jar able to hold 1.5-2L
7. Aluminum foil
8. Filter disc's or cotton wool
9. Glove box or flow hood
5. metal measuring spoon (needed if using peroxide)
6. 3% hydrogen peroxide (optional)
1. Add 50g of MEA into the canning jar, add 1L of water stir till its mostly dissolved, cover the jars opening with the filter disk and screw the outside metal ring back on the jar (if you don't want to use filter disc's just plug the opening with the cotton) now wrap the top with aluminum foil.
2. Place jar into pressure cooker (if you are planing on using peroxide toss the metal spoon in there also) add some water to the cooker and sterilize for 30min at 15psi.
Do not exeed 45min the MEA will start to caramelize and fungi doesn't grow to good on caramelized sugars.
3. Once you can open the pressure cooker again move the jar (and spoon) into your glove box or by your flow hood while still hot.
4. (When using peroxide: once the outside of the jar is manageable but still pretty warm around 120F-140F add 8ml of peroxide and swirl the jar to mix it, be careful not to over agitate the agar and create bubbles)
5. Open your sleeve (keep the sleeve we will need it again) of petri dishes and stack them inside your glove box or in front of the flow hood right side up (I stack mine in tens but it depends on how big your hands are and glove box)
6. Lift the first entire stack by the lid of the bottom dish and slowly pour just enough agar into the dish to cover the bottom. Repeat until all are filled. 1l per 20-30 100ml petri dishes.
7. Stack the completed dishes on top of each other and loosely cover with the sleeve they came in to ensure that they cool evenly this will also minimize condensation on the upper plates.
8. Allow plates to cool overnight
9. Now tape the sleeve shut and store them agar side up in a cool dark place.
now i need to go smoke and then il be back to write up the actual tissue transfer procedure
uromastyx
Well-Known Member
The tissue transfer is a pretty easy process just have to be able to keep things sterile.
Basically what you do is you break the mushroom open lengthwise and then cut a little bit piece out of the inside and place it onto the agar from there it will colonize the agar and you can cut pieces of that and transfer them into other mediums.
Materials:
1. Glove box
2. petri dishes with agar
3. Scalpel
4. Alcohol lamp
5. Alcohol
6. Cotton balls
7. Mushrooms
1. Add all the equipment into your glove box and sterilize everything.
2. Pick a suitable mushroom(s) you want to take one from the cake that colonized the fastest, has the biggest fruits, potency,... Clean all the casing material off of it if there is any stuck to it (best if done away from your work area)
3. Working inside your glove box you want to wipe down the outside of the mushroom with alcohol using a cotton ball
3. Holding the mushroom at its base apply some pressure to it. This should cause the mushroom to split open, now peel the mushroom apart lengthwise through the cap.
4. Sterilize your blade using the alcohol lamp. Cut a small piece of mycelium roughly 3-8mm square from the inside of the mushroom and avoid cutting all the way through to the outside (the inside of the mushroom should be sterile since it has not been in touch with air until you opened it up inside your sterile glove box) Now lightly pierce the fragment with your scalpel and place it in the middle of the agar.
5. Repeat as many times as you like using a new petri dish for each cut and before each cut you should sterilize your blade again.
6. Seal the plates with parafilm, label and date, then place them right side up into your incubator until they grow out then place them upside down.
You should be able to see growth within a few days at first it will be a little fuzz forming as the cells start to divide again but then it should start spreading out. Once colonized you can cut pieces of that and add them to a new medium and grow those. The petri dishes are not meant for long term storage they will get contaminated after a couple of weeks but you can do subculturing agar to agar transfers
All the supplies i get from a website specializing in mycological products they have everything you could want i can post it if someone is interested
Basically what you do is you break the mushroom open lengthwise and then cut a little bit piece out of the inside and place it onto the agar from there it will colonize the agar and you can cut pieces of that and transfer them into other mediums.
Materials:
1. Glove box
2. petri dishes with agar
3. Scalpel
4. Alcohol lamp
5. Alcohol
6. Cotton balls
7. Mushrooms
1. Add all the equipment into your glove box and sterilize everything.
2. Pick a suitable mushroom(s) you want to take one from the cake that colonized the fastest, has the biggest fruits, potency,... Clean all the casing material off of it if there is any stuck to it (best if done away from your work area)
3. Working inside your glove box you want to wipe down the outside of the mushroom with alcohol using a cotton ball
3. Holding the mushroom at its base apply some pressure to it. This should cause the mushroom to split open, now peel the mushroom apart lengthwise through the cap.
4. Sterilize your blade using the alcohol lamp. Cut a small piece of mycelium roughly 3-8mm square from the inside of the mushroom and avoid cutting all the way through to the outside (the inside of the mushroom should be sterile since it has not been in touch with air until you opened it up inside your sterile glove box) Now lightly pierce the fragment with your scalpel and place it in the middle of the agar.
5. Repeat as many times as you like using a new petri dish for each cut and before each cut you should sterilize your blade again.
6. Seal the plates with parafilm, label and date, then place them right side up into your incubator until they grow out then place them upside down.
You should be able to see growth within a few days at first it will be a little fuzz forming as the cells start to divide again but then it should start spreading out. Once colonized you can cut pieces of that and add them to a new medium and grow those. The petri dishes are not meant for long term storage they will get contaminated after a couple of weeks but you can do subculturing agar to agar transfers
All the supplies i get from a website specializing in mycological products they have everything you could want i can post it if someone is interested
Bwpz
Well-Known Member
I'm very interested in that website. Thanks so much for writing those up
Me, being a newbie at this, should I go with the LC or the agar? They said LC isn't good for PF Tek, would this be better?
uromastyx
Well-Known Member
I prefer agar over lc but lc is probably easier for a beginner.
http://www.mycosupply.com/ is the website
http://www.mycosupply.com/ is the website
Bwpz
Well-Known Member
Thanks man, I've repped you before so it won't let me :\ In your experience does LC work for the PF Tek?
uromastyx
Well-Known Member
I don't normally do pf tek anymore and have never tried it with brf before, but i don't see why it should not work. It might take a little longer than normal to be fully colonized but other than that i don't see how there would be any other issues other than contaminants.
Bwpz
Well-Known Member
Makes sense. What is the tek that you use to grow? After this grow I was thinking about doing a bulk grow, but didn't know which would be the best to do.