Stare Sticks: Agar X-Fers Without Flames or Blades

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stareatclouds
star eat clouds?

Registered: 09/29/14
Posts: 10,418

Stare Sticks: Agar X-Fers Without Flames or Blades

8
#22662316 - 12/16/15 09:19 AM (10 years, 8 months ago)

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

Brought to you by THE TRIBE

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)

Extras:

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