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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: Shroombitz]
    #27076166 -

This was incredibly helpful, thank you kind sir. An interesting point about the PC as it cools, and sucking in contams. First time effort: After I opened the PC I tightened the legit GL45 bottle lid with a rubber mit immediately, then let it naturally cool overnight (everything took way longer than expected due to my noob ways). Next morning trying to get it back into microwave I noticed the bottle lid was ON, almost felt fused on. I almost tore my wrist in half trying to open it, I had to get a heavy duty wrench to get it loose again. Was this the vacuum or the cooling shrinkage I wonder? Is this normal? Anyway I've since hacked my way thru the steps and now I'm eagerly awaiting results. Thanks again.:grin:


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"Take it easy dude... But, take it!" -TM

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: MerryTerry]
    #27086177 -

glad yall found it useful, thanks for the kind words :smile:

id love to use glass but it doesnt fit into my workflow, id need to hire a full time dishwasher haha


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C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: c10h12n2o]
    #27117776 -

very useful read. thank you :heart:

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: c10h12n2o]
    #27129538 -

Thank you so useful


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Trying my hand at this cultivating thing.
Thank you so much to this community for the good reads and teks to follow.

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: c10h12n2o]
    #27203202 -

Great post.  Any tips for someone who is transferring MS syringe to agar to isolate from contams but wants as many different strains as possible (looking for clone candidates)?  If I were to inoc a spawn jar with 1.5 cc I'd get a much larger gene pool to choose from.  I just shot up 6 bags with an MSS and 4 were good so there appears to be a low chance of contams in any one inoc loop's worth with this current syringe.  How many drops of inoc can I get away with on a plate before the water creates its own problems?  And do denser clusters of strains make it more difficult to detect contams or do they still stand out?  Thanks!

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: kidcharlemagne]
    #27208755 -

In my opinion, genetics piled on top of each other can make it harder to ID clean cultures.

Also, I believe you are investigating the amount of available genetics in a drop of spore solution. With the average syringe, if you get germination you'll have plenty of genetics to choose from. I would not suggest tripping on trying to use more.


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Masters and slaves hunting clones        How not to isolate (it worked, but)


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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: verum subsequentis]
    #27229501 -

:whathesaid: 100%

I try to use less rather than more

The streaking technique is all about thinning it out, basically a serial dilution on a plate. I usually do it with a print these days, but one drop should work fine if they arent too clumped together

There is TONS of genetic diversity in a tiny bit of spore solution  / print


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C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: c10h12n2o]
    #27375157 -

I hope its okay for me to ask here!:
Where should I transfer off of these plates?
Are they actually good to go to grains now or should I use them as parents for another set?
Thank you and god bless
First dish is a pin transfer from another agar+agar piece from different strain, second dish is a piece from one dish and a piece from another that somehow got broken apart while trying to place it on the dish... I think

Heres some pics of the backs maybe somebody could tell if something hard to see was grown over



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yeah keep telling yourself that's voluntary

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: tututotutut]
    #27380024 -

i dont like transferring to agar or grain when the plates are grown out that far

some people always let em grow out that far before going to grain but personally i like to be able to see what the leading edge looks like (especially for transfers)

what transfer are you on?

the growth looks very unorganized

there are a few patches of good looking myc but at this point everything is very intermingled. you could transfer from those to agar and try to catch it sooner next time, but i wouldnt go to grain or bet that its clean at this point


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C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: c10h12n2o]
    #27380229 -

Thanks! Yeah I had explained in another thread too, somewhere along the way I convinced myself that one must use a full clean dish for successful grain spawning. Completely forgot about spawning with wedges and little pieces.
I'll bounce back lol.
Oh and many of my plates have been in my fridge for months. I was anticipating transferring most of them back to new agar anyway.


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yeah keep telling yourself that's voluntary

Edited by tututotutut (07/09/21 08:38 AM)

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: c10h12n2o]
    #27449145 -

hey m8,

thanks a lot of this wealth of information.

i'm just trying to understand every aspect of this great universe of aseptic work, so therefor: why do you wrap your items in foil before PC? like your scalpels etc. ? what purpose does that serve in the process?
thanks in advance humans

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: incal] * 1
    #27450047 -

happy to help!

i wrap the scalpels in foil envelopes so that i can handle them in open air before unwrapping them in the FH

i usually do lots at a time and use them as needed, storing the wrapped extras in a covered jar until they are needed

for the agar bottles, the foil is an extra precaution, since i transport the agar bottles in open air before they get in the flow. it also helps keep the most important part of the bottle (the top portion that might be above dishes while pouring) sterile. after i get it in the flow, i remove the foil and wipe everything with alcohol before opening

for anything with a filter on it (jars) the foil is just to keep the filters from getting too wet


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

C10's Agar Guide + Tips and Tricks | c10's Flow Hood Build Guide


"Partial knowledge is more triumphant than complete knowledge; it takes things to be simpler than they are, and so makes its theory more popular and convincing."

"Convictions are more dangerous enemies of truth than lies"
― Friedrich Nietzsche

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: c10h12n2o]
    #27527206 -

:headbanger:


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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: c10h12n2o]
    #27571184 -

Lots of great info here.

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: c10h12n2o] * 1
    #27614106 -

I'm a noob and getting started in agar and found this to be extremely helpful.  Thank you!


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- FreeTheFungus

End prohibition now - the war on drugs is worse than the drugs

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: FreeTheFungus]
    #27617388 -

Thanks! I finally understand using a loop and streaking and why you need to flame between the streaks.  Great explanation.

After inoculation, can I let the dishes sit in the SAB for a day before sealing them?  I've done this in the hope that some of the condensation might evaporate -- about 1/3 of my plates have so much moisture that I can't really see the agar surface. (I've been keeping these upside down.)

Would it be too hazardous to quickly absorb the extra moisture on the lid with a piece of sterile cloth or perhaps a bit of toilet tissue?

Finally... I have used about 100 plastic petri dishes and are all contaminated or dried up.  Can I sterilize and reuse these?

To reduce cost and plastic, I've ordered clear ketchup cups and small Ball jelly jars.  After inoculation, should I try to seal these containers -- perhaps with Scotch tape or something that is easier to apply than parafilm? The jelly jars can be tightened down and the little plastic cups are supposedly airtight but.....?

Thanks for such a helpful post!

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: Moe Lawn] * 1
    #27619129 -

I would seal the dishes immediately. The condensation should already be good by the time you inoculate. If you're having issues with that, you should optimize your pouring strategy, e.g., pour when your temperature is lower, keep them stacked when they cool, or place hot coffee mugs on top, etc.

Don't try and wipe off the excess moisture with something. It's possible, but adds an extra layer of risk and your time would be better spot eliminating the condensation in the first place.

Don't use tape. Just buy a roll of Saran/cling wrap and use a sharp knife to cut them into finger-size rolls. Saran works fine as a Parafilm substitute and has the added benefit of not drying out.

Don't store your plates upside down. Plastic petri dishes can't be re-used as the polystyrene melts in a PC. Use Glad Mini Rounds a la Pasty's agar Tek if you're not already. Perfect to use while you get the hang of agar.


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ATTACK OF THE KILLER APE!!!11 | G2G Video | Stare Sticks | Bags in SAB |
DMT + Laser = Reveals Alien Code Commonly Reported In Trips | JavaScript I Use To Make Shroomery Better [New]

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: c10h12n2o]
    #28096299 -

Thank you so much!


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:mushroom2:

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: alexmorgan114]
    #28096325 -

This is an example of a thread that is getting long in years but remains valuable and relevant. A search with a 5 year limit would miss this gem.

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Re: c10's Agar Guide + Tips & Tricks (post #1000) [Re: c10h12n2o]
    #28114543 -

Im going to read this.


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I slit the sheet.
The sheet I slit.
And on that slitted sheet, I sit.

I've always wanted to say that.

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Mushrooms, Mycology and Psychedelics > Mushroom Cultivation
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