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Invisiblestareatclouds
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Stare Sticks: Agar X-Fers Without Flames or Blades * 7
    #22662316 - 12/16/15 09:19 AM (8 years, 1 month ago)

:chief: Stare Sticks :chief:
Easy agar transfers and LC's
without scalpels, needles or flames


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Firstly, transfers are an absolutely essential part of agar work. They allow you to isolate clean growth, keep a desired culture alive for many years, whittle strains down to a monoculture, and so forth.

Despite transfers being such an essential technique for every mycologist to learn, they frustrate me. There’s nothing inherently more difficult about making clean transfers than other aspects of myco-work, I guess I’m just lazy. :lol:

So I tried making transfers to keep varieties alive without the blade or flame.

Stare Sticks:

1. To begin, I recommend isolating clean growth. This should always be the first step when expanding a culture as the cleanliness of an initial MS spore drop is unreliable.



Furthermore, the precision needed to transfer from the leading edge of the healthiest looking mycelium is only available with the tried and true transfer with a sterile blade.

2. Once you’ve isolated clean growth, we’re ready to begin a batch of Stare Sticks.

Note: I’ve nearly always used Pasty Plates for my agar work, and I don’t think standard agar plates will work for this as they’re not tall enough to accommodate our Stare Sticks.

The only difference is that you’ll accompany your plates with a small piece of wood, e.g., Popsicle stick splinter, broken tongue depressor, or toothpick. I’ve successfully used toothpicks and tongue depressors.



3. Break up your chosen piece of wood into small pieces. The length should be just long enough that it can stretch from the bottom corner to the upper rim region of the cup. This allows for the wedge to stay upright during sterilization where your agar liquefies.

Note: You’ll want your agar poured thick enough to cover the bottom corner of your Stare Stick. You want the initial uncovered portion of the wood to be closer to the middle of the plate, not the corners/edges.



4. Once you’ve sterilized your plates, you’re ready for your first (and last) standard transfer. Following sterile procedure, transfer a clean wedge of mycelium as close to the protruding edge of wood as possible. This will allow for quick colonization of the wood.



5. Once your mycelium is climbing the wood, you’re ready to transfer your Stare Stick. In your SAB, prep everything as per usual, although you can forget about your flame and blade. With your plates wiped down thoroughly, loosen each lid enough to be easily separated, and place them both upside down on an iso-soaked paper towel.

Note: It’s helpful to have the Glad Round handle facing you. Simply place a finger on it to help remove the lid easier.

Carefully open your master plate, keeping it as unexposed as possible, and remove your Stare Stick. Once removed, pick up your receiving plate, still upside down, and plant in your colonized piece of wood.

Note: Using a sterilized utensil, such as PC'd tweezers, is probably a safer bet than gloved fingers and iso. I've had success with keeping the container nearly upside down and only using gloves/iso, but you can never be too safe.

Once your stick is secured in the receiving plate, place the lid back on. You’re done.



Note: Depending on how your mycelium has colonized your wood, you may want to place the stick upside down. A tongue depressor, which is wider than it is round, might only colonize the top, making a flip in between transfers optimal.

Additional Thoughts:

  • It is possible to isolate away from satellites and weird growth with this method, but it definitely lacks the precision and flexibility of a standard transfer (hence us beginning with an already transferred and clean growth). However, it is much quicker and has proven reliable as a method of transfer, and you can always use your scalpel on plates inoculated with Stare Sticks down the line.

    I think someone could expand a culture quickly by placing 3-4 toothpicks in a single plate and following standard procedure after.

  • One can also utilize the colonized piece of wood as the transfer vehicle for an LC. I dropped a colonized toothpick into a standard Karo solution, allowed it to sit and germinate for a month, before aspirating into a 60cc syringe via SHIP.



I dropped my LC onto a clean plate. Despite it swirling around everywhere due to excess water, the LC appeared clean after testing on agar. 



  • To ensure your transfer is closer to the center of a plate, I recommend transferring to a larger receiving plate than your initial one. For example, a toothpick from a Pasty Plate going to a ½ pint wide-mouth jar is very accommodating. However, there’s likely no reason you can’t use your sticks as a one-off swab either.

Here’s a quick picture example of the process:



Welp, that’s it. It’s not some revolutionary technique, but I figure it might help people who hate making standard transfers.


Edited by stareatclouds (12/23/15 06:22 PM)


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Invisiblecc2
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: stareatclouds]
    #22662410 - 12/16/15 09:51 AM (8 years, 1 month ago)

nice :thumbup:


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InvisiblePastywhyteMDiscord
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: cc2]
    #22662466 - 12/16/15 10:10 AM (8 years, 1 month ago)

Nice idea there, could also be a good way to increase the lifespan of a plate. Keeps the culture viable without slanting. Will try soon.


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Invisiblestareatclouds
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: Pastywhyte]
    #22662509 - 12/16/15 10:29 AM (8 years, 1 month ago)

I initially started this as a poor man's slant, but I read the small area of the slant tube is what makes it viable for so long, not the wood. Then I realized I could transfer the toothpicks. Either way, I still have a fridge with a bunch of colonized toothpicks for a rainy day. :smile:

Excited to see you try it man. I want it tested by an expert!


Edited by stareatclouds (12/16/15 10:30 AM)


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Offlinetripdawg420
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: stareatclouds]
    #22662537 - 12/16/15 10:35 AM (8 years, 1 month ago)

:yaysugar:


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OfflineMushroom_J
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: tripdawg420]
    #22663066 - 12/16/15 01:03 PM (8 years, 1 month ago)

I'll be trying this.


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Offlineinvitro


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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: Mushroom_J]
    #22663211 - 12/16/15 02:06 PM (8 years, 1 month ago)

Nice one stare, you're a rising star. :mushroom2:


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Invisiblestareatclouds
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: invitro]
    #22663234 - 12/16/15 02:12 PM (8 years, 1 month ago)

A rising stare. Very excited to see people try this as described. I know I can improve the Tek very much with your guys' help.


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Invisibledankington
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: stareatclouds]
    #22663368 - 12/16/15 02:38 PM (8 years, 1 month ago)

This is extremely interesting. Are you actually sterilizing the wood? at all?

On a similar note, wood chopping blocks (I cook) are more contam resistant than plastic ones. No telling, but guaranteed; chop the same thing up on both types and see what you get. Wood breathes, maybe that's why it works?


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Invisiblestareatclouds
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: dankington] * 1
    #22663469 - 12/16/15 03:01 PM (8 years, 1 month ago)

Yes, it's going into my agar plates before the pressure cooker. I sterilize the sticks along with the agar, then inoculate with a standard transfer, and then transfer the wood by hand in a SAB.

I believe wood is used for slants because the mycelium colonizes the divots and lies dormant when refrigerated.

Also, while researching that, I came across an old post where RR and some guy talked about toothpicks or wood being used as the transfer medium itself. No idea's original!


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Invisibleshroominmyroom
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: stareatclouds]
    #22663491 - 12/16/15 03:07 PM (8 years, 1 month ago)

:flyhigh:


Edited by shroominmyroom (12/12/17 04:12 PM)


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Invisibleeatyualive
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: shroominmyroom]
    #22663873 - 12/16/15 04:38 PM (8 years, 1 month ago)

beautiful stare, im proud of you!:rockon:

ill have to try this soon. the blinking gif graphics are a nice touch. and the video is a great example. it looks really easy to transfer.


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Edited by eatyualive (12/16/15 07:49 PM)


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OfflineDetroitMagic
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: stareatclouds]
    #22663933 - 12/16/15 04:51 PM (8 years, 1 month ago)

This is a great addition to the pasty's easy agar plates. I will def be trying this ASAP. Have you noticed it helping the mug climb away from any contams  if you get any? Or do contams grow quickly up the wood as well?

It seems like it would be easy to use a blade to take a bunch of transfers off the stick as well, I know the intent it to skip that step but it might make that step easier as well.


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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: DetroitMagic] * 1
    #22663990 - 12/16/15 05:02 PM (8 years, 1 month ago)

How did I not think of this!! gawd I stare at sterile prewrapped tounge depressors a few times a day.

Ding ding ding no more flaming scalpels for me. EXCELLENT idea props.


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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: newrook]
    #22664399 - 12/16/15 06:17 PM (8 years, 1 month ago)

Kinda loving this idea with toothpicks.  You could get a *little* bit of the precision back by putting in a ring of toothpick pieces.  When the culture grows out, transfer the ones from the areas that look the best.

If you stick em in vertically, the transfer is just a matter of pulling a toothpick and sticking it in center of the new dish, the myc should already be close to the right height to jump right off and grow evenly around, without the unevenness that happens when a wedge goes on its side and myc has to grow around the wedge.

Sounds kinda awesome actually, and would make transfers *so* much easier and more sterile, working in a too-small SAB.


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OfflineSivarted
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: Sivarted]
    #22664416 - 12/16/15 06:22 PM (8 years, 1 month ago)

Ok, obvious mis-think about doing a bunch vertically.  Shit is liquid when it's pcd.

Durr.

I like the idea enough that I wonder if you could get away with making tiny holes in the bottom of the plate, stick half or a third of a toothpick in it, and run em that way.

Agar wouldn't permeate and leak through the wood, would it?


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OfflineSivarted
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: Sivarted]
    #22664422 - 12/16/15 06:24 PM (8 years, 1 month ago)

Hmm.  I think I'm going to try that when I have a chance.  Drill holes, stuff em with toothpicks, put some agar in, and see if I can PC without the agar finding it's way out the bottom


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OfflineSivarted
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: Sivarted]
    #22664479 - 12/16/15 06:36 PM (8 years, 1 month ago)

Or...

If the whole toothpick-plug thing worked out, you wouldn't even need to depend on having enough in there to hope that the sections you wanted grew into one.  Just put three or four around the very outside, and use them to get a scraping to move into the new dish.  You're supposed to transfer a tiny wedge anyway, probably would be easy enough to get a big enough scraping on the end of a toothpick for transferring from wherever you wanted.

I *love* the idea of having presterilized tools inside the dishes themselves.

Sorry, now I'm just spamming the thread with a bunch of random shit, I'll stop.  :laugh:


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OfflineSivarted
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: Sivarted]
    #22664539 - 12/16/15 06:44 PM (8 years, 1 month ago)

Ok one more idea then I swear I'm done until somebody else chimes in:

Scuff up the bottom, put a layer of silicone in, cure it in a PC so the surface is flat and even, then just pour your agar over the top of the silicone and stick the toothpicks into it.


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OfflineMycologist217
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Re: Stare Sticks: Agar X-Fers Without Flames or Blades [Re: Sivarted]
    #22664740 - 12/16/15 07:29 PM (8 years, 1 month ago)

This is great! Cool Tek


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