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PurePleasure
Untrusted bacteria cultivator.


Registered: 08/29/14
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Re: Brf Substitute Tek Progress + Suggestions [Re: Psilosoulful]
#21935138 - 07/12/15 09:13 PM (8 years, 6 months ago) |
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Those jars look great dude. You pulled it off. I would even use the last one. Also just one of those cultures should be enough to inoculate all 7. I would transfer out half of one culture to other dishes and use 1 and a half on your jars and keep the other culture in the fridge for a backup plan
-------------------- If you aren't happy with what you have now, how could you be any happier with more? (Get your SGFC high.)

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Psilosoulful

Registered: 09/05/14
Posts: 7,205
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Re: Brf Substitute Tek Progress + Suggestions [Re: PurePleasure]
#21935222 - 07/12/15 09:34 PM (8 years, 6 months ago) |
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Thanks man!! I didn't expect such good growth either. This BRF paste tek I found was just something to keep me busy until I could get agar supplies. Now I can get growing and work on my sterile procedures in the SAB. I've always used a glovebox up until now with moderate success.
How long do you think cultures can be stored for in the fridge and will they be just as effective when used again?
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PurePleasure
Untrusted bacteria cultivator.


Registered: 08/29/14
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Re: Brf Substitute Tek Progress + Suggestions [Re: Psilosoulful]
#21935585 - 07/12/15 11:28 PM (8 years, 6 months ago) |
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Quote:
Psilosoulful said: Thanks man!! I didn't expect such good growth either. This BRF paste tek I found was just something to keep me busy until I could get agar supplies. Now I can get growing and work on my sterile procedures in the SAB. I've always used a glovebox up until now with moderate success.
How long do you think cultures can be stored for in the fridge and will they be just as effective when used again?
I've used cultures a year old. Trick is is after you take them out the fridge, let them sit out for at least a day or a few days. That will give the myc time to come out of shock and wake back up pretty much. I believe they become a little less effective after time but nothing to really worry about. Just after it wakes up, make a transfer and then cut out the first piece of quickest expansion and drop it into a other plate. Drop it on grains, get mushrooms, and then start back over with spore prints to agar. Then clean it up on agar more. It only takes a few transfers usually to get quick, healthy, rhizmorphic growth. Personally, I let my plates fully colonize, then take the most rhizmorphic piece (usually at the end of the plate) and drop it on agar. It will just continue to produce more and more rhizmorphic growth. Some people do a million transfers and end up with a monoculture which is one set of genetics and produces and acts the same everytime but most monocultures suck. Get good healthy growth on your plate sand go with the flow
-------------------- If you aren't happy with what you have now, how could you be any happier with more? (Get your SGFC high.)

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Psilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
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Re: Brf Substitute Tek Progress + Suggestions [Re: PurePleasure]
#21935678 - 07/12/15 11:58 PM (8 years, 6 months ago) |
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Thanks for the info, it's really helpful I just transfered the brf jar with the most rhizo growth to the seven grain jars. Hopefully all goes well and no jars contaminate. For my first time working in my an SAB , i think I did pretty good, followed all the sterile procedures, but there'e always room for improvement.
I'm gonna store the last two brf paste jars in the fridge for now. I wanna see how these grain jars do before I make any other transfers.
I also have some spore prints from my last grow. I might go ahead and make another batch of brf jars and throw that on there and see if I get clean growth. Soo manny options right now!!
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PurePleasure
Untrusted bacteria cultivator.


Registered: 08/29/14
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Re: Brf Substitute Tek Progress + Suggestions [Re: Psilosoulful]
#21935711 - 07/13/15 12:13 AM (8 years, 6 months ago) |
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Quote:
Psilosoulful said: Thanks for the info, it's really helpful I just transfered the brf jar with the most rhizo growth to the seven grain jars. Hopefully all goes well and no jars contaminate. For my first time working in my an SAB , i think I did pretty good, followed all the sterile procedures, but there'e always room for improvement.
I'm gonna store the last two brf paste jars in the fridge for now. I wanna see how these grain jars do before I make any other transfers.
I also have some spore prints from my last grow. I might go ahead and make another batch of brf jars and throw that on there and see if I get clean growth. Soo manny options right now!! 
The key to sab work is to have a bleach soaked paper towels or towel on the bottom. I do like 50 50 bleach water. Then spray the walls of your sab including the roof with mostly dosh soap and just enough water to where you can spray it, but it doesn't drip. Any contams become stuck to walls for the most part and when they hit the bottom they are neutralized and you have a clean place to set your scalpel or what ever tools you may be using on. I got lazy with my most recent transfers and didn't even take my hands out the sab. I took a straight concentrated bleach paper towel and wiped my blade down in between each transfer and then set the scape down with the bleach rag around the blade covering it. Just make sure there isn't to much bleach on your scalpel when you cut into your plates and after you cut a sectio, try to stab it from the side and catch jt where the uncolonized media is under the mycelium on top. I did 10 ksss plates. I didn't get ONE comtam. Was kinda suprising but I might stick with that instead of going in and out and in and out of your sab. Also wipe down your grain jars with either bleach solution or isobefore putting them in the sab. Be quick because bleach fumes can be strong so try not to mix your bottom towel to loud.
-------------------- If you aren't happy with what you have now, how could you be any happier with more? (Get your SGFC high.)

Edited by PurePleasure (07/13/15 12:15 AM)
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Psilosoulful

Registered: 09/05/14
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Re: Brf Substitute Tek Progress + Suggestions [Re: PurePleasure]
#21935813 - 07/13/15 12:57 AM (8 years, 6 months ago) |
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That sounds like a good approach to take, the contams from outside the SAB have less of a chance of getting inside if you don't take your hands out. Wouldn't making a glove box be the same thing tho? There's so much debate of whether a glove box vs an SAB is better. I always thought that the less airflow there is inside the box with/without gloves would give the best chance of no contams catching itself on exposed plates?
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PurePleasure
Untrusted bacteria cultivator.


Registered: 08/29/14
Posts: 5,893
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Re: Brf Substitute Tek Progress + Suggestions [Re: Psilosoulful]
#21935837 - 07/13/15 01:08 AM (8 years, 6 months ago) |
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I have pvc caps as my arm holes that have a screw on fitting for when ever you need to seal it. I also took a sheet of thin rubber flexible gasket like material and glued it to the inside of my sab on the ends of both caps. The cut horizontal spits in them. So when you go in , you push through the slits and makes a pretty good sield around your long latex loves. I guess you could call it a hybrid  Some may call it overboard, in just call it inenginuity But yes the whole idea of a still air box is... well.. still air
-------------------- If you aren't happy with what you have now, how could you be any happier with more? (Get your SGFC high.)

Edited by PurePleasure (07/13/15 01:10 AM)
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Psilosoulful

Registered: 09/05/14
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Re: Brf Substitute Tek Progress + Suggestions [Re: PurePleasure]
#21935843 - 07/13/15 01:12 AM (8 years, 6 months ago) |
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Sounds cool, I'll look into making those adjustments, I'm just hoping these grain jars do well! Only time will tell
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PurePleasure
Untrusted bacteria cultivator.


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Re: Brf Substitute Tek Progress + Suggestions [Re: Psilosoulful]
#21935858 - 07/13/15 01:22 AM (8 years, 6 months ago) |
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Ingot a feeling they will buddy just be quick with the cut, have your jar come out the pc still in foil and go straight to the sab if you don't wipe them down. Loosen your lids to where you can just pull it right off(one at a time as you go) , drop and seal your jar quick. . Leave it in the sab until your done with ur jars and one there all done and closed, then take them out. Also after you bleach or flame your blade, have the agar or what ever dish loose so you can pull it off with one hand then just set the lid back on top after you cut. If you drop a chunk on the bleach towel just forget about using it. So smoth, steady , and confident wins the race
-------------------- If you aren't happy with what you have now, how could you be any happier with more? (Get your SGFC high.)

Edited by PurePleasure (07/13/15 01:26 AM)
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PurePleasure
Untrusted bacteria cultivator.


Registered: 08/29/14
Posts: 5,893
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Last seen: 11 months, 23 days
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Re: Brf Substitute Tek Progress + Suggestions [Re: PurePleasure]
#21935863 - 07/13/15 01:27 AM (8 years, 6 months ago) |
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Of and the rubber thing i did isn't necessary if you don't want to.
-------------------- If you aren't happy with what you have now, how could you be any happier with more? (Get your SGFC high.)

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Psilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
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Re: Brf Substitute Tek Progress + Suggestions [Re: PurePleasure]
#21936386 - 07/13/15 08:31 AM (8 years, 6 months ago) |
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Yeah I could be faster with cutting from the culture. And I need to make sure I don't hover around the exposed jars either. I'll get better with this in time. I'll definitely look into using the bleach/water spray. This time around, I used a soap/water spray throughout the SAB and wiped down each jar with iso before placing them inside. I also flamed the knife with a cigar lighter each time before making a cut. Hopefully my sterile procedures pay off tho, I hate seeing contams!!
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spacechildo
proletarians rise up



Registered: 01/24/13
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Re: Brf Substitute Tek Progress + Suggestions [Re: Psilosoulful]
#21937210 - 07/13/15 12:54 PM (8 years, 6 months ago) |
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I wouldn't take sab advice from a guy who always has a crapton of bacteria in his grows 
bleaching and believing it "neutralizes" mold spores is just whack. checkout how frank and TL does there sab prep and work.
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Psilosoulful

Registered: 09/05/14
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Re: Brf Substitute Tek Progress + Suggestions [Re: spacechildo]
#21937811 - 07/13/15 03:28 PM (8 years, 6 months ago) |
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I took a look at their teks, really good info for next time I use a SAB. Also I was gonna make a liquid innoculant with one of my brf paste jars. Once I inject the sterile water into the jar and move it around a bit, I'll suck up the mycelium water back into the syringe. Do you think I should store this syringe at room temp or should I leave it in the fridge until I'm ready to use it on grains or another jar?
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mycomaniac1402
Myco Maniac :-)



Registered: 01/25/15
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Re: Brf Substitute Tek Progress + Suggestions [Re: Psilosoulful]
#21937945 - 07/13/15 04:02 PM (8 years, 6 months ago) |
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Id refrigerate because of the BRF. Its already living mycellium so it doesn't need to expand. But before you use , id take it out and let it recover for 24 to 48 hours. I do the same when using a Lc. But thats just me and who knows why I do the things I do lol
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Psilosoulful

Registered: 09/05/14
Posts: 7,205
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Re: Brf Substitute Tek Progress + Suggestions [Re: mycomaniac1402]
#21937998 - 07/13/15 04:13 PM (8 years, 6 months ago) |
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I'll do that for sure, I've read about a lot of people doing the same thing.
Edited by Psilosoulful (07/13/15 04:14 PM)
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spacechildo
proletarians rise up



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Re: Brf Substitute Tek Progress + Suggestions [Re: Psilosoulful]
#21938025 - 07/13/15 04:17 PM (8 years, 6 months ago) |
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I've said it before but I'll say it again you really should take a transfer first.
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Psilosoulful

Registered: 09/05/14
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Re: Brf Substitute Tek Progress + Suggestions [Re: spacechildo]
#21938287 - 07/13/15 05:18 PM (8 years, 6 months ago) |
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Quote:
spacechildo said: I've said it before but I'll say it again you really should take a transfer first.
I will, I still have one more fully colonized jar left. I'm gonna pc more brf paste jars tonight and do some transfers tomorrow!!
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Psilosoulful

Registered: 09/05/14
Posts: 7,205
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Re: Brf Substitute Tek Progress + Suggestions [Re: Psilosoulful]
#21941984 - 07/14/15 01:01 PM (8 years, 6 months ago) |
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Made a nice mycelium filled syringe with one of the brf paste jars. Put it in the fridge just now. 
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mycomaniac1402
Myco Maniac :-)



Registered: 01/25/15
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Re: Brf Substitute Tek Progress + Suggestions [Re: Psilosoulful]
#21942161 - 07/14/15 02:02 PM (8 years, 6 months ago) |
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Looks good man , that mycellium will kick into high gear and colonize quick. Ive made mycellium slurry but never BRF slurry. Was it puffy and lack linear growth? What im asking is was it more difficult to determine good growth on the paste because of the nutes rather than a agar media
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Psilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
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Re: Brf Substitute Tek Progress + Suggestions [Re: mycomaniac1402]
#21942216 - 07/14/15 02:17 PM (8 years, 6 months ago) |
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It didn't isolate the dish, it grew from MS. I have one plate left that I'll use to isolate sections onto more brf jars. But, the mycelium was thick and fluffy all around the plate and there was rhizo growth all around the sides. I'm almost 100% positive it was clean spawn. Anyway, once I make some agar plates, I'll test it out to be sure it's contam free before knocking up more jars with the syringe. 
I already have 7 WBS jars that I transferred a brf wedge to that had similar growth patterns. I'll take the best looking jar of the group, once 100% of course, and use it as a master grain jar. The rest I'll spawn to make my first full size mono 
Now it's time to wait for some growth, I'm gonna have some beer to celebrate lol!!
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