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OfflineKizzle
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Re: Pasty Agar Tek [Re: Inocuole]
    #20760321 - 10/27/14 04:55 PM (9 years, 3 months ago)

I have trouble transferring wedges if the agar is too thin. I just wind up mutilating the whole plate trying to pick it up :lol:


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OfflinePudgy
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Re: Pasty Agar Tek [Re: Kizzle]
    #20760336 - 10/27/14 04:59 PM (9 years, 3 months ago)

Interesting. I'll remake my plates this evening then.

Can I let your agar scrubbie slush cool overnight in my SAB with my pasty plates?


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InvisibleViolet
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Re: Pasty Agar Tek [Re: Pudgy]
    #20760423 - 10/27/14 05:16 PM (9 years, 3 months ago)

I wouldn't suggest it... you gotta swirl or splash that cooling agar up onto the scrubbie.  When it's hot like that, almost all will drip quickly to the bottom!  It must be swirled onto the scrubbie before it gels.  You'll see how it works if you try agar poms a few times.


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InvisibleInocuole
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Re: Pasty Agar Tek [Re: Pudgy]
    #20760428 - 10/27/14 05:17 PM (9 years, 3 months ago)

Ah maybe deep dishes aren't so bad then.  A few of mine are about a cm or more thick.  Just used up the last of them for a couple transfers.  I tried using a guitar string as a loop and it didn't work so well for grabbing mycelium, though I think it might have still worked.  Maybe I'll ask my lady to sacrifice a pair of her tweezers, that does sound effective.  I feel like the tweezer approach would be a lot more like picking shit up and setting it back down, than stabbing it and trying to get it loose.


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InvisibleViolet
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Re: Pasty Agar Tek [Re: Inocuole]
    #20760451 - 10/27/14 05:22 PM (9 years, 3 months ago)

Tweezers cut down into agar as you'd envision a screwdriver doing (by the way, long screwdrivers make great agar tools too!), and can do it from both sides of the intended wedge, then can pick them up and set them down.  The wedge might stick to the tweezers, but if you stick the bottom of the wedge to the new agar it will hold to it more and let the tweezers go.


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The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
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InvisibleInocuole
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Re: Pasty Agar Tek [Re: Violet]
    #20760464 - 10/27/14 05:26 PM (9 years, 3 months ago)

Any old tweezers will do?  Or are you using super long tweezers that are more meant for lab-related stuff?


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InvisibleViolet
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Re: Pasty Agar Tek [Re: Inocuole]
    #20760505 - 10/27/14 05:36 PM (9 years, 3 months ago)

Well if they're short and small they're probably more annoying than they're worth.  Hygiene tweezers might not cut it, depends. The ones I have are lady's hygiene tweezers, maybe four inches long if that, like this.. They work for me.  But I've oft considered ordering some long ones like these.  The ones with the wide tip like women's tweezers would cut down into agar the best though.  We can make it work with anything but the sharp tip ones.
If buying tweezers, ones like this should be the very shortest worth putting money to.


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Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
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InvisibleInocuole
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Re: Pasty Agar Tek [Re: Violet]
    #20760528 - 10/27/14 05:42 PM (9 years, 3 months ago)

Cool she just said I can use the slant-tip ones that she doesn't like.  She seems to prefer the needle ones for whatever ungodly reason.

Too bad I just did my transfers and am out of clean plates.  I wonder if it would be worth it to sterilize the tweezers in an empty pasty plate with a little bit of water to create some steam, and then just open it up in the SAB already sterile so you don't have to pre-flame it for the first transfer.

Also, does anyone know if senescence is triggered by total metabolization over time (general aging/amount of sub consumed) or by number of reproductive cycles?  For instance, if I've transferred a grain to agar, it pinned, transferred that pin to agar, and repeated that like 2-3 more times, so that I'm on the third or fourth generation of agar pin clones, will I experience problems when I finally spawn to grain?


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InvisiblePastywhyteMDiscord
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Re: Pasty Agar Tek [Re: Inocuole]
    #20760571 - 10/27/14 05:54 PM (9 years, 3 months ago)

Its a by product of cell division and age. A full cycle into a fruit body will require a lot of cell divisions which is why we usually only clone from ms. I would not want to transfer pins more than twice. Usually when I get an invitro pin I transfer it, isolate to master plates. Then once I have tested the genetics I take the master plate that did best and transfer that to a slant for storage. The rest of the plate with be transferred to several others and they will be used to inoculate.


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OfflinePudgy
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Re: Pasty Agar Tek [Re: Violet]
    #20760769 - 10/27/14 06:32 PM (9 years, 3 months ago)

Quote:

Violet said:
I wouldn't suggest it... you gotta swirl or splash that cooling agar up onto the scrubbie.  When it's hot like that, almost all will drip quickly to the bottom!  It must be swirled onto the scrubbie before it gels.  You'll see how it works if you try agar poms a few times.




I'm getting ready to make the mixture. I figured, if I splash the scrubbie and get it coated it should be okay to hang out until morning.

That isn't right? :frown:


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InvisibleInocuole
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Re: Pasty Agar Tek [Re: Pastywhyte]
    #20760775 - 10/27/14 06:33 PM (9 years, 3 months ago)

Cool, I already have some put to grain, but I have another batch of transfers that I'm going to want to inoculate from as well.  I'll have to check into making a slant.


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InvisibleViolet
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Re: Pasty Agar Tek [Re: Pastywhyte]
    #20764862 - 10/28/14 05:32 PM (9 years, 3 months ago)

Quote:

Inocuole said:
Cool she just said I can use the slant-tip ones that she doesn't like.  She seems to prefer the needle ones for whatever ungodly reason.
I wonder if it would be worth it to sterilize the tweezers in an empty pasty plate with a little bit of water to create some steam, and then just open it up in the SAB already sterile so you don't have to pre-flame it for the first transfer.



I wrap mine slightly loose in foil up to an inch or less of the back.  With good sterile procedure, or especially in sterile airflow, you're pretty much only sterilizing after the first transfer for the consideration of cross-contamination.


Quote:

Inocuole said:
Also, does anyone know if senescence is triggered by total metabolization over time (general aging/amount of sub consumed) or by number of reproductive cycles?  For instance, if I've transferred a grain to agar, it pinned, transferred that pin to agar, and repeated that like 2-3 more times, so that I'm on the third or fourth generation of agar pin clones, will I experience problems when I finally spawn to grain?



Well, it depends on how you're taking the pins.  I would suggest doing it just ONE time.  And there is a very good way of going about this.... 
Quote:

Pastywhyte said:
Its a by product of cell division and age. A full cycle into a fruit body will require a lot of cell divisions which is why we usually only clone from ms. I would not want to transfer pins more than twice. Usually when I get an invitro pin I transfer it, isolate to master plates. Then once I have tested the genetics I take the master plate that did best and transfer that to a slant for storage. The rest of the plate with be transferred to several others and they will be used to inoculate.



Bingo.


Just for a nice example using no-pour agar dishes, I have these examples from my no-pour tek with screw-tops like I use for my grow, my most important tech http://www.shroomery.org/forums/showflat.php/Number/19035259#19035259 which just had its first birthday 2 days ago!

Quote:

Pins will take a few days to fuzz-up and start growing across the agar.


It's best to transfer pins as early as possible into maturity.

Some sterile pins transferred quite late:

At a certain point they may even continue to mature and open sporulating caps. We definitely don't want that for a clone dish.

These dishes were all started with small pins:


Growth is much more ideal; these cultures are and will continue to be more healthy and vigorous.

From there it's a snap to isolate the fastest & strongest of the mycelium that grows from the pin and start a small test batch!




--------------------
Intentionally or not, here in mushcult we are purveyors of love culture and enlightenment movement. Let's try to act like it!

PODS TEK - Growing Invitro with BRF/verm or Grass Seed containers
The simplest, quickest, safest tek!  For beginners, culturers, lazy people, stealth lovers, contam haters, and alternative seekers!
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OfflineFreeWorldOrder
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Re: Pastywhyte's Easy Agar Tek [Re: Pastywhyte]
    #20767243 - 10/29/14 09:39 AM (9 years, 3 months ago)

Just made my first ones and they look good. I took someones advice (can't remember where I read it). And pot a small folded pieces of paper towel over my micron luer-lok filter then the normal foil (used microns on my lids for GE)This helps soak up condensation and appears to have worked well. Thanks to Patsy and everyone else! I believe I am on my way to some successful cultures.


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OfflinePudgy
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Re: Pastywhyte's Easy Agar Tek [Re: FreeWorldOrder]
    #20767527 - 10/29/14 11:17 AM (9 years, 3 months ago)

I made some thin plates. Probably, 3-4mm. The myc is growing alright. I'll definitely know more tomorrow. 3rd transfer. I'm not sure how my Violet's BRF Gravy Grow is doing, the gravy sludge boiled all over the walls so I can't see ****.


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OfflinePudgy
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Re: Pastywhyte's Easy Agar Tek [Re: FreeWorldOrder]
    #20767534 - 10/29/14 11:18 AM (9 years, 3 months ago)

Quote:

FreeWorldOrder said:
Just made my first ones and they look good. I took someones advice (can't remember where I read it). And pot a small folded pieces of paper towel over my micron luer-lok filter then the normal foil (used microns on my lids for GE)This helps soak up condensation and appears to have worked well. Thanks to Patsy and everyone else! I believe I am on my way to some successful cultures.




Congrats!

Can you elaborate more on your anti-condensation technique? I'm very interested. Though wiping with sterilized napkin has worked well -- if I can skip that I will.


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Offlinekmetric
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Re: Pastywhyte's Easy Agar Tek [Re: Pudgy]
    #20774399 - 10/30/14 08:06 PM (9 years, 2 months ago)

Just noticed something that looked like myc growing up the sides of my pasty plate..weird. Don't think it's cobweb, took a couple of weeks to get to that point.


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InvisiblePastywhyteMDiscord
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Re: Pastywhyte's Easy Agar Tek [Re: kmetric]
    #20774884 - 10/30/14 09:46 PM (9 years, 2 months ago)

Pics?


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OfflinePudgy
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Re: Pastywhyte's Easy Agar Tek [Re: Pastywhyte]
    #20776214 - 10/31/14 08:21 AM (9 years, 2 months ago)

If you're like me... An untalented savage for a scientist then somehow you splattered mycelium all over the place inside your plate! One of my plates fell out of my hands and the wedge bounced off of every wall and when trying to get it back to the center, it rubbed all over the agar surface....

FUCK!!! I was trying to isolate that shit LOL! Now it's just silly.


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InvisiblePastywhyteMDiscord
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Re: Pastywhyte's Easy Agar Tek [Re: Pudgy]
    #20776375 - 10/31/14 09:38 AM (9 years, 2 months ago)

It happens to the best of us, just keep at it, practice is key :thumbup:


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InvisibleInocuole
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Re: Pastywhyte's Easy Agar Tek [Re: Pastywhyte]
    #20777114 - 10/31/14 01:10 PM (9 years, 2 months ago)

Dude my last batch of transfers is fucking littered with satellites.  I don't know wtf I'm doing wrong, maybe I need to douse my gloves in iso better first, maybe I need to get a spray bottle with 50/50 iso/water and mist the pasty plates before I open them?  I've been turning off the ac for about an hour, spraying the walls of the SAB, using damn near elbow-length gloves, and keeping the dishes up toward the top of the SAB while I open them so things can only come from the limited space above.  Using a mask too, to diffuse my stank breath a bit.  I got little bacterial dots (I presume that's what they are) all over like 3/4 plates I just did the other day.

So it's either coming from the tool I'm using, wind currents that shouldn't exist, or my own body somehow.  Could it really be that I just need to take a shower first, or what?


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