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OfflineBLINKfan420
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Registered: 07/06/12
Posts: 1,078
Last seen: 5 months, 1 day
Saved My Golden Teachers!! PICs =)
    #18880912 - 09/23/13 02:40 PM (10 years, 4 months ago)

So after going through months of time and work to acquire and fruit a GT syringe that I nearly lost.. here are the little babies that will hopefully drop tons of spores so I can continue there existence.




I had 12 jars fail off my Golden Teacher MS syringe that I made. (Bad procedure I guess).  I double inoculated 4 more.  3 of the double 'noced jars failed as well. I had the last cc of spores I had left, and squirted them ALL into the last jar. The 16th (last) jar made it to full colonization but a week after birth contam'ed with trichoderma.  I cut the cake in half to save it, and then it came back.. I cut it in half again (hence the wedge) so that less than a quarter of the cake was left.

At that point I figured I lost my GTs for good.. and I couldnt order/much less find another syringe.

a few days later.. to my surprise.  Theyre saved

I will be much more careful making a new MS syringe this time!


--------------------



All my pictures are drawn from imagination.  I don't even know what a mushroom is.


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Offlinecmori3
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Re: Saved My Golden Teachers!! PICs =) [Re: BLINKfan420]
    #18881019 - 09/23/13 03:18 PM (10 years, 4 months ago)

Agar, young grasshopper.



Apparently a good technique is to do a few agar transfers, transfer some agar into a liquid culture, and then after colonization squirt some of said liquid culture onto agar - it will grow out and if no bacteria grows from the inoculation point then you know you are all good. Important side note - bacteria may grow on the plate in general due to air contamination. Try and keep the plate as unexposed to air as possible so that this level of confusion does not enter into the equation. Once you know you have a clean liquid culture, you can inoculate the same as if you had a MS syringe. It should colonize considerably faster too.


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OfflineSockadin
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Re: Saved My Golden Teachers!! PICs =) [Re: cmori3]
    #18881150 - 09/23/13 03:55 PM (10 years, 4 months ago)

You have strived to save something special! Nice Job.

I wish you the best, and I am curious as to your MS Printing/needle work.


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OfflineBLINKfan420
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Registered: 07/06/12
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Re: Saved My Golden Teachers!! PICs =) [Re: Sockadin]
    #18883072 - 09/23/13 11:52 PM (10 years, 4 months ago)

I am now a botany major and mycology is just as interesting as plants, so I am super excited to try agar.  I need to look into a good place to buy agar dishes or a good recipe to whip up my own. 

In the past I take a thick piece of foil and sterilize with isopropyl alcohol about three times over (just in case)

Then do the same to the inside of a glass mug.

I cut the cap as soon as its open and is flat and carefully place it on the foil then set the mug over it.

After 24hr I have a THICK black (purple w.e) print.

I take a brand new syringe out of the package..

suck up some boiling water

let sit for 30 min or so..

From there I isopropyl sterlize a shot glass..

Scrape spores with flame sterlized razor blade into shot glass.

Squirt pasteurized water into shot glass..

Stir..

and suck up the spores.

wait 24hrs and its good to go.

**Im sure there are better ways


--------------------



All my pictures are drawn from imagination.  I don't even know what a mushroom is.


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OfflineMush4Brains
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Registered: 07/31/13
Posts: 4,419
Last seen: 9 years, 2 months
Re: Saved My Golden Teachers!! PICs =) [Re: BLINKfan420]
    #18883109 - 09/24/13 12:05 AM (10 years, 4 months ago)

As long as it is all done in a still air box you should be good.


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Offlinemyplague240


Registered: 03/17/13
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Re: Saved My Golden Teachers!! PICs =) [Re: Mush4Brains]
    #18883178 - 09/24/13 12:40 AM (10 years, 4 months ago)

:whathesaid:


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InvisiblePastywhyteMDiscord
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Registered: 09/15/12
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Re: Saved My Golden Teachers!! PICs =) [Re: myplague240]
    #18883577 - 09/24/13 05:59 AM (10 years, 4 months ago)

Prints taken from fruits that were fruited in open air are not sterile. Isopropyl will only sanitize not sterilize. Pasteurized water? I am amazed that you even got this far. Start researching agar and G2G. This will allow you to perpetuate not only your variety, but also any individual strains that you may happen to like :thumbup:


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OfflineBLINKfan420
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Re: Saved My Golden Teachers!! PICs =) [Re: Pastywhyte]
    #18884279 - 09/24/13 10:30 AM (10 years, 4 months ago)

Quote:

   Prints taken from fruits that were fruited in open air are not sterile. 




So there is a close to 100%/100% sterile way to take a print?

I'm a noob so sorry if I don't use the correct terms.    Sterile water is what I guess I meant by pasteurized.  But boiled water that kept at that temp. for however long and then is rapidly cooled would by definition be pasteurized.. so I thought


--------------------



All my pictures are drawn from imagination.  I don't even know what a mushroom is.


Edited by BLINKfan420 (09/24/13 10:33 AM)


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Offlinempd
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Re: Saved My Golden Teachers!! PICs =) [Re: BLINKfan420]
    #18884459 - 09/24/13 11:13 AM (10 years, 4 months ago)

That would be the definition of pasteurizing.


--------------------
There is no truer calling for mankind than that of true conservatism.


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InvisiblePastywhyteMDiscord
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Re: Saved My Golden Teachers!! PICs =) [Re: mpd]
    #18884826 - 09/24/13 12:58 PM (10 years, 4 months ago)

To make a sterile print will be difficult. You could try fruiting invitro but you still might encounter issues getting the fruit out. I do my printingin my SAB in as clean of conditions as I can muster, and often have no contams show up when I go to agar. That being said I would not consider my prints sterile. If you want to be self sufficient then I would look into agar. That way you don't need the print to be sterile in order to use it. Also keep in mind that ISO does not sterilize, it sanitizes.


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OfflineBLINKfan420
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Registered: 07/06/12
Posts: 1,078
Last seen: 5 months, 1 day
Re: Saved My Golden Teachers!! PICs =) [Re: Pastywhyte]
    #18885759 - 09/24/13 04:44 PM (10 years, 4 months ago)

If I'm printing onto foil whats the best way to keep it sterile??

I've made several prints and several syringes from those prints with no problem. I just turn off the A/C.  Bomb the bathroom with Lysol and wipe down with ISO.  Put on a mask and gloves. Then do the procedure I mention above.  All the syringes have been good too the last drop and nice and dark as well... Except the GT one.  Omg what a disaster....


--------------------



All my pictures are drawn from imagination.  I don't even know what a mushroom is.


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Offlinemyplague240


Registered: 03/17/13
Posts: 127
Last seen: 10 years, 2 months
Re: Saved My Golden Teachers!! PICs =) [Re: BLINKfan420]
    #18886464 - 09/24/13 07:57 PM (10 years, 4 months ago)

BLINKfan420, you don't have to go to that far of extremes. Best way to make a print IMO is to take a half pint jar that has a self healing injection port (Made out of RTV silicon) Clean it out before hand with 91% alcohol, wipe off a pair of scissors... then take a fresh clean needle and stab it into the top of the mushroom cap... take the scissors and cut under the cap and place it in the jar and put the lid back on.

It also helps if you have a marble in the jar before hand so when you put water in for the syringe it's easier to break up the spores stuck to the bottom.

Ps: Doing any kind of mycology work in the bathroom is just an all around bad idea. A bathroom is a breading ground for mold and bacteria.

:thumbup:


Edited by myplague240 (09/24/13 08:02 PM)


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OfflineElement1
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Registered: 04/08/13
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Re: Saved My Golden Teachers!! PICs =) [Re: myplague240]
    #18887405 - 09/24/13 11:51 PM (10 years, 4 months ago)

nice job, but you should check out g2g (grain to grain) transfers. you basically just take a very tiny sample of myc tissue (cut from the center of the cake is the cleanest and best place to sample) and use that to start a LC or just place your bit of tissue under the dry verm barrier on a PF cake. no need to worry about contaminated syringes! (of course its always good to have spores so you can start again if everything possible goes south)

otherwise good luck!


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InvisiblePastywhyteMDiscord
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Re: Saved My Golden Teachers!! PICs =) [Re: Element1]
    #18887979 - 09/25/13 06:16 AM (10 years, 4 months ago)

:smbfacepalm:


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