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Offlineflaw
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Registered: 01/30/99
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Transplanting mycellium bring new life
    #452071 - 11/08/01 08:54 PM (15 years, 10 months ago)

Ok, I posted this in standard mushroom cultivation, didn't get much of a bite at it.

I recently laid plans for a cultivation project. I wasn't too happy about sterility and my fears were realised with almost 100% contamination. Never mind, first time I've ever had significant contamination.

Anyway I have two jars which are doing fine, except one thing - they are too dry, dry enough for the mycellium to not really grow very fast. (I did not allow for the significant water loss experienced in microwave sterilisation).

I propose to use the already grown mycellium to make some new syringes, so i can restart the jars.

So obviously I am seeking guidance in which way is the most effective in using the spawn to create syringes. I'm fairly experienced at growing so give me your wacky idea's if you will also :)

thanks

Incidentally, I plan to publish a microwave sterilisation tek on here at some stage because I have been able to get almost 100% success rate using it in the past.



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Offlinehoneyroasted
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Re: Transplanting mycellium bring new life [Re: flaw]
    #452129 - 11/08/01 09:37 PM (15 years, 10 months ago)

an oven, some tweezers, some isopropyl alcohol, and some sterile jars of substrate. keep everything clean and just transfer mycelium from the slowly colonizing jars to the fresh ones.


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Offlineflaw
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Re: Transplanting mycellium bring new life [Re: honeyroasted]
    #452151 - 11/08/01 09:48 PM (15 years, 10 months ago)

I would prefer to make syringes so I could start more than just a few jars but thanks for your input.


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OfflineElektrolurch
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Re: Transplanting mycellium bring new life [Re: flaw]
    #452451 - 11/09/01 02:46 AM (15 years, 10 months ago)

Then go over agar and produce new syringes from the colonized petri dish.

Elektrolurch


--------------------
"For all the time spent in that room
The doll's house, darkness, old perfume
And fairy stories held me high on
Clouds of sunlight floating by.", Pink Floyd '67


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Invisiblealtarego
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Registered: 10/25/01
Posts: 130
Re: Transplanting mycellium bring new life [Re: flaw]
    #452638 - 11/09/01 10:40 AM (15 years, 10 months ago)

If you had contamination problems in the other jars, you might also have it in these two. Not all contaminants are visible. Trying to go from jars back to syringes only invites problems.

If you know how to do agar work, then I would suggest taking fully colonized, individual grains and placing them (with flame-sterilized tweezers) onto plates, or better yet, peroxidated agar plates, growing them out and going to jars from there.

At least that way you will be sure the culture is really clean.

Or you could let the jars colonize, fruit them and make new spore syringes from the fruits.

- AE


Edited by altarego (11/09/01 11:36 AM)


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Offlineflaw
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Re: Transplanting mycellium bring new life [Re: altarego]
    #453000 - 11/09/01 04:09 PM (15 years, 10 months ago)

Altarego,

I am aware of the fact that the jars themselves could be contaminated. I doubt that for two reasons, one: every other jar contaminated in less than 3 days after innoculation (massive contamination, with full colonisation shortly after) and secondly I believe that the reason the jars are stalling is that they are far too dry.

I may be wrong, certainly I've experienced what you're talking about, the shock of finding out that a jar is contaminated when all looks fine.

I might go back to using spores, just i can't guarantee myself that the prints I have taken are not the source of the contamination. I have taken successful prints before with no problems and I certainly didn't do anything 'less' in taking those prints.



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OfflineGroove_N
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Registered: 09/22/01
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Re: Transplanting mycellium bring new life [Re: flaw]
    #453036 - 11/09/01 04:41 PM (15 years, 10 months ago)

Is the substrate in the jars PF cake method, or grain?
I have lots of sucess using the microwave and PF cakes.


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Invisiblealtarego
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Posts: 130
Re: Transplanting mycellium bring new life [Re: flaw]
    #453044 - 11/09/01 04:47 PM (15 years, 10 months ago)

Flaw -

I know what you mean. If you think the spores might be contaminated, then your best bet is to fruit the jars you think are ok and take new prints and/or clones. Then you can be sure that the cultures are clean.

Peroxidated agar is another good bet.

Best of luck
AE


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Offlineflaw
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Re: Transplanting mycellium bring new life [Re: altarego]
    #453683 - 11/10/01 04:56 AM (15 years, 10 months ago)

Groove, its a PF method and I too have had much success. This time it didn't work out unfortunately.

I don't know whether I can let these jars grow to fruiting, I think they are all but stalled completely. I might consider doing something drastic like injecting sterile water into the cakes.



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OfflineGroove_N
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Registered: 09/22/01
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Re: Transplanting mycellium bring new life [Re: flaw]
    #456171 - 11/12/01 06:00 PM (15 years, 10 months ago)

Flaw:
One problem I have encountered when Microwaving is drying out (as you mentioned above). This can be eased by adding more water to your substrate before microwaving. What is the magic ratio??? Another problem I encountered when using the microwave early on is the you need to have some air exchange for the mycelium to grow, other wise it stalls. Seems like this may be the case you have. If you have a dry verm layer on top, I would recomend some kind of air exchange. If you are going to make a new micorwave Tek, I would be more that willing to help you out.


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