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OfflineMadHatR
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Registered: 10/13/03
Posts: 278
Last seen: 3 years, 3 months
First time with Agar, will this work for no touch spore to LC process?
    #26894832 - 08/23/20 06:32 AM (3 years, 5 months ago)

Ok, so it's been about 15 years since I have practiced mycology. With plenty of time on my hands, I thought I would pick it up again. Back when I was practicing, it was commonplace to knock up a LC jar with a spore syringe, wait a week, make LC syringes then knock up some quart jars and put them in a incubator for a few weeks. During that time only a few people had access to a flow-hood and Priority Mail envelopes were your best defense against contamination.

Fast forward to today. I have read up on new tek's and now agar is king, flow-hoods are commonplace and isolating is the key to faster growth which will overcome contamination.

Until I get more comfortable with my sterile environment and get a flow-hood then I would like to limit chances of contamination. Following that idea I was thinking of combining a few of Pasty's Teks and would like to know if it would work.

With what I have access to I was thinking of getting some Ziploc 4 oz Mini Round Twist 'N Loc Containers. They are screw on so from what I was reading I should be able to cook up some MEA, and spoon in a layer of agar in each one. I currently don't have any injection ports or Whatman syringe filters, so for the first batch I was thinking of making a SHIP with some Permatex High-Temp RTV Silicone and for GE I can PC them with the lid 1/4 turn loose.

The idea I had would be to PC the Mini Rounds with the modified lid with a SHIP. Once it cools down I can open the PC, tighten the lids, remove the foil and place the Rounds with a layer of agar in my SAB. I would then be able to take my spore syringe, flame sterilize the needle and inject a small amount into the sealed agar round. Remove, re-flame and repeat.

By the time they have colonized then I could use a 14 Gauge needle to push through the SHIP and take a biopsy of the desired growth, pulling it up in a syringe of PC distilled water. I could then take the needle with the biopsy out and inject it into a 1/2 pint jar that has a Whatman syringe filer, injection port and agar on the bottom.

Wait another week or so until it has colonized nicely, then inject 120cc of water, breaking up the colonized part into the water. I would then be able to make tons of LC syringes.

From that point I can knock up a bunch of quart jars of rye berries that also have LC lids on them.

By doing this everything is done inside sterilized jars, from spore to spawn.

Will this work? Have I missed or overlooked anything? It is basically Pasty's Easy Agar Tek and EzLC Tek. The difference being instead of dropping agar wedges in and cutting wedges out for transfer everything is done through a SHIP.


--------------------
Never Judge a man until you walk a mile in his shoes, because then you are a mile away and you have his shoes.


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