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OfflineCloneufc
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Cloneufc's easy Penis Envy spore tek! * 4
    #14085370 - 03/08/11 06:33 AM (12 years, 10 months ago)

Most of the time PE will not drop spores at all. This tek will teach you how to extract spore's from the Penis Envy strain, the easy way. You wont ever have to worry about PE spores again. This tek has worked wonders for me with PE swabs. Swiping PE to agar has produced mycelium only once for me. With this tek I have gotten swabs to start mycelium on agar 99% of the time. If your gonna use this tek for swabs just substitute the cap for the swab. This tek does not require sterility. Clean but not sterile. Also don't dump the spores into an LC, bad idea.

First pick a PE cap that has a purplish tint to it's gills. (Many PE are sterile)


Next you will need a syringe and needle. Fill the syringe with 5cc of distilled water. The distilled water does not have to be sterile. Now squirt a couple of cc's between the gills of the cap, while sucking the water back into the syringe. Make sure to use the flat part of the needle or you will have a hard time sucking the water back up. Keep doing this until the water is purple in color.




Now grab a piece of foil and shape it like you would a print. After that is done take the foil and shape it into a cup shape. Take the syringe and squirt the 5cc of spore water onto the foil. The purpose of shaping the foil into a cup shape is so that the spore water will stay in without leaking all over. Now place the print out in the open and let it dry for 1 day.


After all the water has evaporated, your PE print will look like this. I use an inoculation loop to scrap the spores together on foil. Now you can use your inoculation loop to transfer the spores to agar.


Edited by Cloneufc (03/08/11 06:51 AM)


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Offlinebbt0009
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Re: Cloneufc's easy Penis Envy spore tek! [Re: Cloneufc]
    #14085409 - 03/08/11 07:02 AM (12 years, 10 months ago)

I like where your heads at but why didn't you just keep the water in the syringe and shoot up a jar??

:winning: <- I had to use this


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Offlinebbt0009
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Re: Cloneufc's easy Penis Envy spore tek! [Re: bbt0009]
    #14085412 - 03/08/11 07:03 AM (12 years, 10 months ago)

My point here being that instead of storing a print you can just store the syringe in the fridge


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OfflineCloneufc
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Re: Cloneufc's easy Penis Envy spore tek! [Re: bbt0009]
    #14085603 - 03/08/11 08:18 AM (12 years, 10 months ago)

Because every single time I have done it that way, there is bacteria in the syringe. It's probably coming from the gills. Bacteria can't live in a dry environment. This is the best way. I'm not saying it can't be done that way. I write these teks to eliminated the painful steps that lead to failure. Hence  I'm not a fan of spores and liquid mixes. Dry spores and agar are much more successful. I have been doing it this way for years. I have around 7 years of experience and I have a good idea of what works and what doesn't. Dry it out and transfer the spores to agar. Works every time. As a bonus, once it's on agar you can pick the best sectors. You can't do that knocking up a jar with a ms syringe. My advice is if your gonna use a syringe, use one where the PE printed and make a syringe from that.


Edited by Cloneufc (03/08/11 08:23 AM)


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Offlinedietrich
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Re: Cloneufc's easy Penis Envy spore tek! [Re: Cloneufc]
    #14086249 - 03/08/11 11:54 AM (12 years, 10 months ago)

Doesn't leaving the print out in the open to evaporate like that leave it open to a high contamination risk?


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OfflineGonkulator
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Re: Cloneufc's easy Penis Envy spore tek! [Re: dietrich]
    #14086316 - 03/08/11 12:11 PM (12 years, 10 months ago)

That's why he's taking it to agar. To clean up into a good culture.


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Invisiblebiologys
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Re: Cloneufc's easy Penis Envy spore tek! [Re: Gonkulator]
    #14086418 - 03/08/11 12:32 PM (12 years, 10 months ago)

once you start working with agar, you'll never do spores to grain again, you'll always put on agar to be sure you clean culture. and to start isolating.


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Invisiblejokefox
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Re: Cloneufc's easy Penis Envy spore tek! [Re: biologys]
    #14086480 - 03/08/11 12:44 PM (12 years, 10 months ago)

thats not a bad idea at all


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OfflineCloneufc
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Re: Cloneufc's easy Penis Envy spore tek! [Re: dietrich]
    #14086544 - 03/08/11 12:55 PM (12 years, 10 months ago)

No. Dry prints and spores don't need to be sterile when transferring to agar. That's the beauty of prints. There's far too many new people on here making it tougher on themselves.


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OfflineTaesian
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Re: Cloneufc's easy Penis Envy spore tek! [Re: Cloneufc]
    #14086589 - 03/08/11 01:03 PM (12 years, 10 months ago)

Never used agar before, but have a question.

Lets say there's a contam hooked on a spore.

When you throw it on agar, and if that contam starts growing out, you can just cut away a good sector, transfer, and wah-la? Is that what you mean by cleaning out a culture?

So no matter what you can get a clean something out of any print?


Danke schon.


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Invisiblebiologys
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Re: Cloneufc's easy Penis Envy spore tek! [Re: Taesian]
    #14086615 - 03/08/11 01:07 PM (12 years, 10 months ago)

Quote:

Taesian said:
Never used agar before, but have a question.

Lets say there's a contam hooked on a spore.

When you throw it on agar, and if that contam starts growing out, you can just cut away a good sector, transfer, and wah-la? Is that what you mean by cleaning out a culture?

So no matter what you can get a clean something out of any print?


Danke schon.




you are correct

sometimes you get a dish that you cant salvage anything from, thats why you make multiple dishes of it, so you can find one to salvage and get a clean culture from.


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Offlinegornyhuy
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Re: Cloneufc's easy Penis Envy spore tek! [Re: biologys]
    #14087593 - 03/08/11 04:20 PM (12 years, 10 months ago)

I prefer swabs, and have never had any trouble getting PE spores to germinate on agar from my swabs and from others' swabs sent to me from shroomery folks.  :shrug:


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Edited by gornyhuy (03/08/11 04:37 PM)


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OfflineCloneufc
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Re: Cloneufc's easy Penis Envy spore tek! [Re: gornyhuy]
    #14096771 - 03/10/11 08:50 AM (12 years, 10 months ago)

I don't have a problem germinating swabs if I cut a piece of the swab and put it on agar or if I put the whole swab on agar. I do have problems when I try just streaking the swab to agar, no germination.


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Offlinegornyhuy
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Re: Cloneufc's easy Penis Envy spore tek! [Re: Cloneufc]
    #14099753 - 03/10/11 06:33 PM (12 years, 10 months ago)

I hold the swab above the center of the dish (in a glovebox with very sterile procedures) and then use a flame-sterilized scalpel blade to scrape along the side/top of the swab in the direction of the agar... Almost like a flint and steel type of motion.  This has never failed to get a TON of myc germinating in multiple spots on the dish. 

I have tried various other approaches like touching the swab to the agar, using a inoculation loop, and so on, but none of them were very reliable.


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