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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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Re: Pastywhyte's Easy Agar Tek [Re: Inocuole]
#20777223 - 10/31/14 01:48 PM (9 years, 3 months ago) |
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I always take a shower first. How long do you wait for the air to settle when bringing flamed tools back in the box?.I go 20 seconds at least. Also you should wipe the plates down with iso before they go in the box.
Transferring from bacteria can be hard, it could just be riding along. Look up agar sandwich, is quite effective.
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 9,959
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Re: Pastywhyte's Easy Agar Tek [Re: Inocuole]
#20777251 - 10/31/14 01:59 PM (9 years, 3 months ago) |
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I'm just noticing growth in one of the 7 cups I did 5 days ago (seems slow?) and it looks clean so far.
Quote:
Also you should wipe the plates down with iso before they go in the box.
So I'll be taking them out of the PC and then wiping them down? Just the outside or what? None of this was mentioned in the original Tek. If this is important, any chance you can update OP with explanation?
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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No leave em in the foil and put em in the box. But once a plate has grown out and is ready to transfer, the outside should be carefully wiped (I assume it was taken out of the SAB at some point).
The tek was written just with the intent of providing someone with a clean plate. But I should do a writeup on procedure.
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Re: Pastywhyte's Easy Agar Tek [Re: Pastywhyte]
#20777301 - 10/31/14 02:19 PM (9 years, 3 months ago) |
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That makes sense. I don't think the bacteria is riding along because it's always a satellite pretty far from the inoculation point. The transfers themselves seem clean every time, the mycelium itself isn't facing any obstacles at all from what I can tell. Though, it does seem to be vaguely the same type of bacteria each time.
That may just indicate that it's coming from me each time though.
Whatever it is it doesn't seem to have a foothold in the grain jars I transferred to. I imagine the grains are not QUITE as great a medium for it as agar though.
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Edmunter
Mr



Registered: 05/01/13
Posts: 5,699
Last seen: 1 month, 12 days
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Re: Pastywhyte's Easy Agar Tek [Re: Inocuole]
#20777360 - 10/31/14 02:46 PM (9 years, 3 months ago) |
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Since i changed my SAB

I have had much better success.
I plan what im going to do in my head b4 doing it so I have no doubts. When opening a colonised culture I wipe it down b4 putting in the SAB, wipe it down b4 use and open the lid upside down wiping the sides b4 tilting for the job at hand. Tams can get caught under the catches I should imagine.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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Re: Pastywhyte's Easy Agar Tek [Re: Inocuole]
#20777361 - 10/31/14 02:46 PM (9 years, 3 months ago) |
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That's true, agar is a perfect medium however, you do need to figure out the vector. Pics would help.
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 9,959
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Re: Pastywhyte's Easy Agar Tek [Re: Pastywhyte]
#20777419 - 10/31/14 03:12 PM (9 years, 3 months ago) |
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Quote:
Pastywhyte said: No leave em in the foil and put em in the box. But once a plate has grown out and is ready to transfer, the outside should be carefully wiped (I assume it was taken out of the SAB at some point).
The tek was written just with the intent of providing someone with a clean plate. But I should do a writeup on procedure.
Ah, okay. Yeah, the procedure would be an awesome addition to this already awesome Tek. I PC'd my agar, put it in SAB from fridge, inoculated, and put on top of my fridge in an open box. I assume this is fine, I just need to wipe down the outsides with iso. Got it.
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Re: Pastywhyte's Easy Agar Tek [Re: Pastywhyte]
#20777521 - 10/31/14 03:51 PM (9 years, 3 months ago) |
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Quote:
Pastywhyte said: That's true, agar is a perfect medium however, you do need to figure out the vector. Pics would help.
Eh, I dunno if pics are gonna help as much as a video of my procedure would, but I'm not really feeling up to either. It doesn't really matter what it is or what it looks like, it's not mycelium and it's on my plates in small little spots that grow much slower than mycelium.
All I need to know is that I need to improve my technique. And get some more iso.
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 9,959
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Re: Pastywhyte's Easy Agar Tek [Re: Inocuole]
#20777701 - 10/31/14 04:40 PM (9 years, 3 months ago) |
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A video is super easy to make. Just hit record and set it down on your SAB.
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Yeah see that's not the hard part. I know exactly how to make a video, it could even be a decent one. Problem is I don't have anything to transfer, and I don't feel like doing it today, especially since it'd have the potential to be an embarrassment, given that I already have vectors that I know need working on.
Would rather get some practice in. Once I'm particularly confident I'll compile all my methods with some documents and video or what-have-you as a return back to the community.
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Edmunter
Mr



Registered: 05/01/13
Posts: 5,699
Last seen: 1 month, 12 days
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Re: Pastywhyte's Easy Agar Tek [Re: Inocuole]
#20779365 - 11/01/14 03:55 AM (9 years, 3 months ago) |
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It would be interesting to see. I might do this to up my game.
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Inocuole
Scalpel of Evil's Bane



Registered: 11/21/11
Posts: 24,863
Loc: ★
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Re: Pastywhyte's Easy Agar Tek [Re: Edmunter]
#20780101 - 11/01/14 12:11 PM (9 years, 3 months ago) |
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Yeah that's the point really. I have most of the essential knowledge down and I know I could just pull any old regular joe off the street and teach them to grow like a bad ass but I don't always follow the best procedures because I am, personally, a lazy fuck. I do know the best procedures, I just don't always live up to them. Once I can set a little better example I'll be glad to start pumping out resources back out into the community. I just want some pics/vids to show for it and that comes with the proof, which comes with time.
The ratio of good resources to bad resources on this site still isn't optimal and that's why dudes like Pasty/Frank/SBJ doing the good work is such an important thing
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Pudgy
Fail-2-Win

Registered: 09/30/14
Posts: 657
Last seen: 8 years, 9 months
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Re: Pasty Agar Tek [Re: Violet]
#20780382 - 11/01/14 01:36 PM (9 years, 3 months ago) |
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Quote:
Violet said: Extra thin agar can possibly only support thin "rhizo" growth. I myself, as well as others I've seen, have mistaken growth on thin or nutrient-weak agar for great rhizomorph. I no longer care so much about impressive-looking rhizo. My best cultures hardly do it, and I only see it on weak nutrition anyway which I pretty much never use. So it's best to have at least a little bit more than just the minimum required for a working surface area.
Thick agar is very beneficial for storing a container in refrigeration long-term.
If you have problems transferring deep wedges, try poking into the depth sideways with the scalpel after cutting and removing the wedge, or using a second sterile tool (tweezers, my favorite. I usually only use tweezers now).
Two questions:
IF you don't care about rhizo -- what makes your other cultures your best cultures?
I went ahead and spooned about 3-4 mm on the bottom of three plates and my growth is good so far.
MY second question is, why would thick agar be better for refrigeration if the myc is on the top surface and never eats down into the agar?
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,890
Loc: Milky way
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Re: Pasty Agar Tek [Re: Pudgy]
#20780392 - 11/01/14 01:38 PM (9 years, 3 months ago) |
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Quote:
I no longer care so much about impressive-looking rhizo
QFT.
Everyone is obsessed with rhizo. Really beats me why, you don't know how good your culture is until you fruit it.
Quote:
IF you don't care about rhizo -- what makes your other cultures your best cultures?
their performance
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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Thick agar is better for refridgeration cause it won't dry out as fast. Same principle as a slant.
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Pudgy
Fail-2-Win

Registered: 09/30/14
Posts: 657
Last seen: 8 years, 9 months
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Thanks gents!
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 9,959
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Re: Pasty Agar Tek [Re: Pudgy]
#20782555 - 11/02/14 01:33 AM (9 years, 3 months ago) |
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This is about 8 days after inoculation and the only plate with any growth. I used the very, very end (like 1/4 CC left) of a syringe so I'm thinking there weren't many active spores left?

On the plus side, it looks clean, so that's nice. The other 6 are probably bunk and should be tossed?
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Edmunter
Mr



Registered: 05/01/13
Posts: 5,699
Last seen: 1 month, 12 days
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Ive used 3-4 mil in my new plates and the myc growing from my new transfers is very thin. Normal?
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,811
Loc: Canada
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Re: Pasty Agar Tek [Re: Edmunter]
#20782916 - 11/02/14 07:00 AM (9 years, 3 months ago) |
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Yep that looks alright. Hang on to the others if you can you never know. But I would take transfers from that one in a few days.
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stareatclouds
star eat clouds?



Registered: 09/29/14
Posts: 9,959
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How many transfers am I taking from it? And am I cutting out the pieces with the X-acto and dropping them into new plates?
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