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InvisibleBooNanZa
Microbiologist
Registered: 07/05/05
Posts: 16
Mini Monotub Grow and Agar question (Pics)
    #11231878 - 10/12/09 08:50 AM (14 years, 5 months ago)

After several weeks of hard work, I finally got some pins from my mini-monotub. Here are the stats:

1. Cambodian Strain
2. Popcorn Spawn
3. Spawned to straight Coir (substrate)
4. Cased with 60/40 Vermiculite/Coir (out of peat moss)
5. Misted by hand with small fan running near tub for FAE

Here are some pictures from yesterday:


And some pictures from today:


Now here is my question: I just recently obtained some petri dishes and PDA (potato dextrose agar) and have been planning on doing some agar/isolate work.

Now should I:
1. Take a small growing pin from a large cluster and attempt to grow it on agar and start the isolation from there?

OR

2. Use MS Spore syringe and THEN start isolation from there?

I feel like #1 would be easier, since I havent really found a good way to get spore solution onto agar dishes without too much liquid getting on the plate itself. BUT, I still have to make/pour these plates, so would these pins be too far grown to put onto agar by the time the plates cooled? Would I just need a small piece of stem and place directly on agar? If anyone has any experience please let me know, I am really excited about working with agar for the first time.

Here is picture of pins clump I would out onto agar:


THANKS FOR ALL YOUR HELP! A great successful grow that couldn't have been accomplished without you guys!

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Offlinelibertaire
liberator
Male


Registered: 08/06/08
Posts: 4,204
Last seen: 3 years, 8 months
Re: Mini Monotub Grow and Agar question (Pics) [Re: BooNanZa]
    #11231893 - 10/12/09 08:53 AM (14 years, 5 months ago)

You can put the tissue of a full grown mushie on agar.  I think it would actually be wiser to do, because with a pin, you're not certain what it's end traits will be, but with a full grown mushroom, you can select the biggest and healthiest looking one that appears.  Nice job btw!

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InvisibleRonPaulVerm
Piss Christ
Male

Registered: 07/21/08
Posts: 1,072
Re: Mini Monotub Grow and Agar question (Pics) [Re: BooNanZa]
    #11231900 - 10/12/09 08:56 AM (14 years, 5 months ago)

for agar you don't need much tissue...
And too much liquid is not good.
Haven't had any experience with this amount but I've heard one drop is good enough.
Goodluck, have fun!


--------------------
Spawn I Have: Blewitt, Reishi, Maitake, Chicken of the Woods, Parasol, Shaggy Mane, King, Blue and Gold Oyster, Shiitake, King Stropharia, Lions Mane, Almond Portabello, Elm Oyster, Phoenix Oyster, Nameko, Enoki, White Beech,

Spawn I Want: Corcyceps Sp., Cauliflower Mushrooms, Agaricus species (Portobello), Pink Oyster, Piopinno, ...anything else you might have


Primitive Skills                                     

Any information I've posted is only related to fictional purposes. I do not advocate growing any illegal mushrooms 
...I DO advocate learning about
Law Enforcement Against Prohibition

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InvisibleBooNanZa
Microbiologist
Registered: 07/05/05
Posts: 16
Re: Mini Monotub Grow and Agar question (Pics) [Re: RonPaulVerm]
    #11231950 - 10/12/09 09:13 AM (14 years, 5 months ago)

Quote:

libertaire said:
You can put the tissue of a full grown mushie on agar.  I think it would actually be wiser to do, because with a pin, you're not certain what it's end traits will be, but with a full grown mushroom, you can select the biggest and healthiest looking one that appears.  Nice job btw!




That definitely makes sense, since you are looking for the best fruit. But then I also heard that immature pins are good because they are rapidly dividing and so are good for cloning/agar work. But I'm not rally sure? So I can just use the full grown mushie in case my plates don't cool before they mature? Would this be a better option than using spore solution? THANKS for your response, I really appreciate it.

Quote:

RonPaulVerm said:
for agar you don't need much tissue...
And too much liquid is not good.
Haven't had any experience with this amount but I've heard one drop is good enough.
Goodluck, have fun!




So would you suggest using tissue from the mushrooms, or trying to use the MS spore solution? I can't think of a good way to transfer without getting too much liquid with the spore syringe, but would this give me a better isolation. Maybe squirt into sterile shotglass and then swab plate with loop? Anyways, what route do you suggest I take? THANKS for your reply as well, any information is greatly appreciated.

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Offlinelibertaire
liberator
Male


Registered: 08/06/08
Posts: 4,204
Last seen: 3 years, 8 months
Re: Mini Monotub Grow and Agar question (Pics) [Re: BooNanZa]
    #11232049 - 10/12/09 09:42 AM (14 years, 5 months ago)

I would definitely pick tissue over spore solution, because with spore solution, you'll have several germination points on the agar, since there will be several sets of spores that meet in several different areas, resulting in several substrains, which I suppose could be good practice if you're looking to gain experience with isolating sectors, but if you want to just see what it's like to get something going on agar, or to save the genetics of a specific specimen that you like the genetics of, tissue culture would be my choice.  Honestly, one's not better than the other, they're just different, but if you have fresh specimens, I would take advantage of that fact and isolate/clone some genetics.  I mean, hell, if you have enough plates to do so, why not do both methods?

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InvisibleRonPaulVerm
Piss Christ
Male

Registered: 07/21/08
Posts: 1,072
Re: Mini Monotub Grow and Agar question (Pics) [Re: libertaire]
    #11232741 - 10/12/09 12:03 PM (14 years, 5 months ago)

^^^everything he said :thumbup::thumbup::thumbup::thumbup:
do tissue and solution!
tissue is prac. for isolation and it's faster (if you already have a fruit)
p.s. i like your idea about swabbing from a glass. it's like what people do with swabbing from spore prints. But if you can get 1 drop to come out of your syringe you're probably fine


--------------------
Spawn I Have: Blewitt, Reishi, Maitake, Chicken of the Woods, Parasol, Shaggy Mane, King, Blue and Gold Oyster, Shiitake, King Stropharia, Lions Mane, Almond Portabello, Elm Oyster, Phoenix Oyster, Nameko, Enoki, White Beech,

Spawn I Want: Corcyceps Sp., Cauliflower Mushrooms, Agaricus species (Portobello), Pink Oyster, Piopinno, ...anything else you might have


Primitive Skills                                     

Any information I've posted is only related to fictional purposes. I do not advocate growing any illegal mushrooms 
...I DO advocate learning about
Law Enforcement Against Prohibition

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OfflineMigraine
Expanding Life
 User Gallery

Registered: 09/21/09
Posts: 158
Loc: Alabama
Last seen: 4 months, 19 days
Re: Mini Monotub Grow and Agar question (Pics) [Re: RonPaulVerm]
    #11232889 - 10/12/09 12:29 PM (14 years, 5 months ago)

Don't work with pins.

Wait until they fully fruit.

Take a mushroom that has the characteristics you want, (size, potency, etc), and perform a clone to agar process.

If you have never cloned before, look at RR's videos - he has an excellent one on cloning.

Your resulting agar culture will be that of the exact mushroom you liked.

Good Luck.

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