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Invisibleredcrow

Registered: 03/25/22
Posts: 384
PF-tek to agar
    #27816160 - 06/12/22 02:05 PM (2 years, 7 months ago)

I have a couple PF-tek jars that were inoculated from a syringe of Pink Buffalo back on 5/26. They've shown strong rhizomorphic grown and are nearly ready to birth. Since the mycelium in these jars look so ropy and aggressive, I wonder if it would be worthwhile to transfer a small piece to agar.

I thought I'd carefully tease out as clean a piece of rhizome as I can get. I'd expect to do multiple transfers to get a good clean plate, but at least I'd be starting from well identified material.

Or would this be a fool's errand?

Here's one of the jars in question



Edited by redcrow (06/12/22 02:17 PM)

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Offlinesan pedro guy
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Registered: 10/22/17
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Re: PF-tek to agar [Re: redcrow]
    #27816169 - 06/12/22 02:11 PM (2 years, 7 months ago)

I had some cakes that looked good but I fucked something up and they never fruited, no contam visible so I took a piece from the center, put on agar and it was clean after 2-3 transfers. I droppes it to some wheat and I plan on spawning today.

As far as I know myc is myc, namean?


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Noob Grow Along 2022

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Offlinesonoramo
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Registered: 02/27/19
Posts: 950
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Re: PF-tek to agar [Re: redcrow]
    #27816219 - 06/12/22 02:48 PM (2 years, 7 months ago)

Quote:

redcrow said:
I have a couple PF-tek jars that were inoculated from a syringe of Pink Buffalo back on 5/26. They've shown strong rhizomorphic grown and are nearly ready to birth. Since the mycelium in these jars look so ropy and aggressive, I wonder if it would be worthwhile to transfer a small piece to agar.

I thought I'd carefully tease out as clean a piece of rhizome as I can get. I'd expect to do multiple transfers to get a good clean plate, but at least I'd be starting from well identified material...





Do you still have the syringe? I'd suggest buy some long dispensing needles (or blunt needles). At least 16ga (smaller numbers means bigger gauge, so 12, 14, or 16). Make sure they are long enough to reach the mycelium you want. You can get these from Amazon for next-day delivery, like these. Be sure to sterilize the syringes, plungers and needles by putting them in boiling water for 5 minutes, pull them out with chopsticks or tweezers. Then assemble the syringes with needles. Flame the needle tips and sanitize the jar top before you stick anything in through your inoculation holes.

You should have about 1ml of air in the syringe before you put the needle in the jar. That way if you get a mycelium plug in the needle, you'll still have some air in the syringe to push it out. Stick the needle into your favorite mycelial area, pull back on the plunger, then use the syringe to inoculate a plate. If the mycelium is stuck in the needle, it may splat out when you inoculate the agar. Just make sure the little plug lands on the agar rather than the plate sides or cover.

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Invisibleredcrow

Registered: 03/25/22
Posts: 384
Re: PF-tek to agar [Re: redcrow]
    #27816253 - 06/12/22 03:09 PM (2 years, 7 months ago)

Since I'll be doing this when I birth the  cakes and I'll be in front of an FFU, the whole cake will be exposed. So I should be able to dissect a piece of mycelia off using a sterilized scalpel and forceps.

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OfflineDoctorBenway
NatAlchemist

Registered: 06/08/22
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Last seen: 1 year, 7 months
Re: PF-tek to agar [Re: sonoramo] * 1
    #27816674 - 06/12/22 07:42 PM (2 years, 7 months ago)

I'm a low budget kinda guy no expensive fancy things here, what you are suggesting reminds me of a technique I use to make clones lol... I'll let a cake go long enough it starts pinning in the jar then make some clean unused lc and pull some into a clean syringe then poke into a fruit in the jar inject a lil lc into it and pull back then put into the lc jar and let it do its thing. I've had great success doing it that way versus tissue transfer in SAB which has failed me way too many times.


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Offlinesonoramo
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Re: PF-tek to agar [Re: DoctorBenway]
    #27817321 - 06/13/22 09:40 AM (2 years, 7 months ago)

Quote:

DoctorBenway said:
I'm a low budget kinda guy no expensive fancy things here, what you are suggesting reminds me of a technique I use to make clones lol... I'll let a cake go long enough it starts pinning in the jar then make some clean unused lc and pull some into a clean syringe then poke into a fruit in the jar inject a lil lc into it and pull back then put into the lc jar and let it do its thing. I've had great success doing it that way versus tissue transfer in SAB which has failed me way too many times.




I've done that sort of thing too. If I'm going to agar though, I'd rather use air as the "fluid" rather than broth. Otherwise the broth on the agar makes it look irregular, which makes it harder to see bacterial growths starting.

My goal is to never use an SAB or any kind of HEPA rig again, ever. Nurses do that all the time when they stick needles in you, and it doesn't cause "contamination." That's motivating me to come up with ways that do all transfers from anything-to-anything with syringes or swabs.

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OfflineDoctorBenway
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Registered: 06/08/22
Posts: 95
Last seen: 1 year, 7 months
Re: PF-tek to agar [Re: sonoramo]
    #27818860 - 06/14/22 05:20 AM (2 years, 7 months ago)

My first dabbling in this hobby was bout 25 years back, friend helped me setup a small closet with antichamber and seperated shelf storage chamber all sealed in heavy guage plastic. Was working with agar and a strain of copelandia that was active in the mycelia. Even working naked freshly showered with a swimmers cap on charcoal filter mask and decontam the antichamber before opening the shelf and tote with plates id have on average 70% success in clean transfers, which i think was pretty good. These days im working in a semi sterile environmemt and feel like id have a lil bit more issues working with agar again and the lc method ive been using seems to work very good for me as ive not had any issues with consistant contam in ages. If it works, work it. If not adjust and keeping working til it works right?

Edited by DoctorBenway (06/14/22 05:22 AM)

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Offlinesonoramo
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Registered: 02/27/19
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Re: PF-tek to agar [Re: DoctorBenway]
    #27818961 - 06/14/22 08:03 AM (2 years, 7 months ago)

Quote:

DoctorBenway said:
My first dabbling in this hobby was bout 25 years back, ...




I got started even earlier, a few years after Oss and Oeric's book came out. I used a flow-hood in an electronics assembly clean room. Still, nearly everything I did got contaminated, and getting even a couple of small grows direct from mason jars was a struggle. Maybe I should have partnered with a biologist instead of chemists and electrical engineers!

Quote:

Even working naked freshly showered with a swimmers cap on charcoal filter mask and decontam the antichamber before opening the shelf and tote with plates...




That would make a great video. Too bad smartphones weren't yet a thing back in the day!

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OfflineDoctorBenway
NatAlchemist

Registered: 06/08/22
Posts: 95
Last seen: 1 year, 7 months
Re: PF-tek to agar [Re: sonoramo]
    #27819994 - 06/14/22 09:05 PM (2 years, 7 months ago)

Haha wouldn't be surprised if my ex has Polaroid snapshots, she thought my antics were quite entertaining. Good times


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