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Amazon Shop: Agar, Autoclave, Laminar Flow Hood

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OfflineLefty
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Registered: 05/23/04
Posts: 2
Last seen: 13 years, 7 days
? re: LB-agar, also seeking the mycologist aka Erliquin
    #2719197 - 05/23/04 06:36 PM (13 years, 1 month ago)

Hi,

So I've decided to take the plunge and try growing some P. cubensis, from a spore print of a Thai strain I picked up locally. I have access to an autoclave, a flow hood, and other goodies, but I have a question regarding agar medium. The plates I have immediate access to are LB plates, which contain...

10g Bacto-tryptone
5g Bacto-yeast extract
10g NaCl
7.5g Bacto-Agar

in 1L of water. So today I threw a few mg of spores on each one, taped them up, and now I'm going to see what happens... does anyone have any suggestions as to the suitability of this medium for Psilocybe?

'Nuther question... is temperature critical, or does it just speed the process of colonization? I can keep them at 20C no problem and I am not in a big hurry.

Okay one more... I am looking to get in touch with a fellow who also goes by "Erliquin" on another site, we used to both be mods there, but that's all I'm gonna say... Erli if you are still around drop me a PM, I have some interesting news for ya....


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Offlinesuboriginal
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Registered: 05/06/04
Posts: 278
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Re: ? re: LB-agar, also seeking the mycologist aka Erliquin [Re: Lefty]
    #2719485 - 05/23/04 08:08 PM (13 years, 1 month ago)

20c should be fine to get the mycelia happening (is for P. subaerugs anyways), though I have never grown cubes... I would personally try and fortify the agar with some potato/ rice/ sugar broth (boil some chopped up potatoes (skin on) and rice, with some sugar... the amounts are not that crucial... I simmer it all together (in bottled or double-distilled water) for abomut 1/2 hour, to promote sterility... The problem will be mixing the broth through your agar without contaminating it... for this I just tip a bit of the broth into the plates, and microwave it (on defrost) until it's all liquid, then let it set... has worked a treat to grow mycelia but maybe I've just been lucky... I'm no expert... you mentioned you have autoclave/ fume hood access etc... why not simply make up some potato/rice/sugar agar, autoclave it, and pour some of your own plates under the hood ? Plenty of good agar recipes can be found here with more precise amounts, but frankly I've found mycelia to be very forgiving, as long as the plates are inoculated sterily (is that a word?!?)... Hope this helps... good luck.


--------------------
Peace, love and organic brown rice...


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Invisiblemicro
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Registered: 05/09/03
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Re: ? re: LB-agar, also seeking the mycologist aka Erliquin [Re: Lefty]
    #2719596 - 05/23/04 08:24 PM (13 years, 1 month ago)

LB is for bacteria, not mycelium. Bacteria will have an advantage on LB plates -- esp. w/ t/ salt concentration and C:N ratio. It would be a lot better to use something like PDY. Other than that, why would you leave them at 20c? That is really cold and will inhibit the mycelial growth which is already slow to begin with. It would be a lot better to incubate them at room temp. Again, though, you have a large chance of bacteria taking over, since it can grow a lot faster. It will work, though, if you are completely sterile (however you will put environmental stress on the substrain.)

--
Micro


--------------------
Any research paper or book for free
(Avatar is Maxxy, a character by Mizzyam, RIP)


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OfflineLefty
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Registered: 05/23/04
Posts: 2
Last seen: 13 years, 7 days
Re: ? re: LB-agar, also seeking the mycologist aka Erliquin [Re: micro]
    #2722939 - 05/24/04 03:09 PM (13 years, 1 month ago)

Wow, good info, thanks a bunch!

It sounds like the potato starch plates are worth the trouble, I had LB plates already poured so I thought I'd throw some spores at them... if it works, how long should it take to see visible growth? micro thanks for the clarification re: LB, the plates are at room temperature, which probably averages ~20C over any given 24h period.


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