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OfflineForresterM
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: deadmandave]
    #19061142 - 10/30/13 11:32 PM (10 years, 3 months ago)

Quote:

deadmandave said:
This is a weird thought i've had but since i no longer use a glove box i havent tried it.

My idea is to put a small slit in the side of the gb, just big enough to slip the tip of your scalpel through, then on the other side put your lantern which you can then flame sterilize without moving your arms in and out of the box.

Might be counter productive because at that point you'd have a flame moving air around a small hole in the still air box.. :shrug: but i would much rather flame sterilize than trust alcohol before doing agar work.




Who was it... I think Malicom does something similar to that and says it works well.  I think he slides the box just off the edge of the table and sticks it out there to flame, but I can't remember exactly.

I've never flamed a scalpel blade, I just use a new one each time.  I've got a box of 100 and I don't do that much agar work though...


--------------------
Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here.
-------------------

Have some medicinal mushrooms and want to get the most out of them?  Try this double extraction method.


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Offlinedeadmandave
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: Forrester]
    #19061994 - 10/31/13 07:03 AM (10 years, 3 months ago)

Quote:

Forrester said:
I've never flamed a scalpel blade, I just use a new one each time.  I've got a box of 100 and I don't do that much agar work though...




:lol: I might be a bit of a cheapskate but..

I bought 6 scalpel bladed 3 years ago and still i am on number three or something. They last a really long time even though they can get ugly and nasty looking. I'll use a sharp new one if im doing some detailed work but that is a rare occassion. Just flame the f*ck out of it between each and every use i.e. between grain jars, between agar plates, between inoculations.

btw, have ya tried putting extra extra water in your p.c. when cooking agar plates? maybe the extended cooling time would keep condensation from being so pronounced :confused:


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Offlinesocraticd
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: Forrester]
    #19062623 - 10/31/13 10:28 AM (10 years, 3 months ago)

Quote:

Forrester said:
Ugh, condensation issues... having the same problem lately.  I really, really, really wish I had a flow hood.




Well quite honestly this seems to work REALLY well for me, and it does stand to reason that it should be fine.  If your GB can be completely sealed, as mine can (I'll put up pics tonight, promise), there won'tbe any introduction of new contams into the box once your pour work is done.  At that point, the plates are covered/closed and the possibility of contams finding their way in to a box with very little moving air, through the gaps in the petri covers, should be VERY small. 

Obviously there has to be SOME airflow in there to evaporate the condensation, but my belief is that its minimal enough to risk for the reward of condensation free plates.  I'll gladly make an extra couple plates when pouring a batch to aaccount for the MAYBE one per sleeve that gets contaminated.


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Offlinesocraticd
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: deadmandave]
    #19062649 - 10/31/13 10:33 AM (10 years, 3 months ago)

Quote:

deadmandave said:
This is a weird thought i've had but since i no longer use a glove box i havent tried it.

My idea is to put a small slit in the side of the gb, just big enough to slip the tip of your scalpel through, then on the other side put your lantern which you can then flame sterilize without moving your arms in and out of the box.

Might be counter productive because at that point you'd have a flame moving air around a small hole in the still air box.. :shrug: but i would much rather flame sterilize than trust alcohol before doing agar work.




See, I could be crazy, but my thought is that I've seen a LOT of good anecdotal evidence that 70% iso works VERY well.  And if that's to be believed, and I can male an initial assumption that I'm pulling material from a plate that isn't showing obvious contamination already, I'mless llikely to pick up outside air and introduce NEW contaminants by keeping the holes prettyuch plugged up with my arms. 

Time will tell, however, since I'll be doing a lot more agar work in the coming months and will keep experimenting to see what ACTUALLY works versus the pure theory of what should and shouldn't.  Agar and plates are cheap enough that I don't care about losing them to gain information.


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Invisibleliamtheloser
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: socraticd]
    #19063013 - 10/31/13 11:59 AM (10 years, 3 months ago)

Pouring hot agar will cause a LOT of air movement in a still air box,  even a sealed one.  I tried this several times with agar,  each time I poured in a glove box or still air box I had more contams than not.  Once I switched to prepouring plates and then sterilizing them I have yet to get a contaminated plate. I also don't have much problem with condensation, but I don't know why.

Edit: I let my pc slowly cool with the petris inside, usually the top one on the stack has some condensation but the others don't. If you insulate them they will probably stop condensing.


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


Edited by liamtheloser (10/31/13 12:08 PM)


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OfflineMolimo
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: liamtheloser]
    #19063245 - 10/31/13 12:49 PM (10 years, 3 months ago)

I've had only problems with doing agar in a SAB. I started doing my clones with BRF in pint jars and have yet to have a contamination (I've only done about 15 clones). What I do is the standard small amount of BRF + water paste at the bottom of the jars, PC for 10-15 min, and throw my cutting in. After the BRF colonizes, I pour sterile WBS on top of the BRF and let that colonize. Then I can just do grain to grain to quart jars and not mess with cutting up agar slices and sterilizing scalpels. Maybe not the best way, but it works for me! :grin:


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Offlinesocraticd
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: liamtheloser]
    #19063783 - 10/31/13 02:38 PM (10 years, 3 months ago)

Quote:

liamtheloser said:
Pouring hot agar will cause a LOT of air movement in a still air box,  even a sealed one.  I tried this several times with agar,  each time I poured in a glove box or still air box I had more contams than not.  Once I switched to prepouring plates and then sterilizing them I have yet to get a contaminated plate. I also don't have much problem with condensation, but I don't know why.

Edit: I let my pc slowly cool with the petris inside, usually the top one on the stack has some condensation but the others don't. If you insulate them they will probably stop condensing.





Obviously it would cause a lot of air movement.  But the pouring step I have had MAYBE one contam out of probably 100 plates.  Its transferring plates that have been solidified and cooled for a significantpportion of time that has led me to contam.  So far the pour and cool has been a pretty clean procedure for me inside my SAB. 

Are you using glass petri's?  I havent tried pre-pouring and sterilizing in my plastic pre-sterilized dishes yet, but I'm not optimistic about them not melting, from what I've heard.


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Invisibleliamtheloser
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: socraticd]
    #19064051 - 10/31/13 03:27 PM (10 years, 3 months ago)

You have to use glass or polypropylene.

I use polypropylene because they're cheap and reusable. They are around 25 bucks for 100, but they're smaller too. I think I'm using 15x60mm


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


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Offlinedrake89
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: liamtheloser]
    #19064351 - 10/31/13 04:21 PM (10 years, 3 months ago)

the drawback to prepouring plates, in my experience, is that you have to do a real thin pour.  with a nice normal pour you will end up with agar all over the place.


--------------------
Fiery Fungi (like us on faeboo)


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Invisibleliamtheloser
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: drake89]
    #19064729 - 10/31/13 05:19 PM (10 years, 3 months ago)

Quote:

drake89 said:
the drawback to prepouring plates, in my experience, is that you have to do a real thin pour.  with a nice normal pour you will end up with agar all over the place.




Never had a problem,  what did you run into?


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Offlinesocraticd
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: liamtheloser]
    #19072528 - 11/01/13 10:10 PM (10 years, 3 months ago)

Finally come pictures I've been promising you guys. 


New still air/glove box.  LOVE this thing, since it can be used and completely sealed back up afterwards which has really helped with the plates, as i mentioned above.  Though, I may upgrade to a larger tub, since this one does get a little crowded when I have a bunch of things in it.



The flask I bought for pouring plates.  WAY easier than the beer growler i was attempting to use before.




The 11 plates of reishi I expanded from the fully colonized plate that i grew out from the wedge that Forrester was kind enough to donate to my projects.  I owe you some wedges once I get up and running with some more species, bud!




A closeup of one of the plates.  This strain moves QUICKLY.  As you can see, this was a small wedge transferred three days ago and it;s expanded to about the size of a quarter, at this point.  Looking healthy!



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OfflineForresterM
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: socraticd]
    #19072629 - 11/01/13 10:34 PM (10 years, 3 months ago)

Man, you're looking like an agar pro there!  Your SAB kicks the crap out of mine.  Gettin' jelly here!! :lol:


--------------------
Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here.
-------------------

Have some medicinal mushrooms and want to get the most out of them?  Try this double extraction method.


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Invisibleliamtheloser
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: Forrester]
    #19073523 - 11/02/13 03:05 AM (10 years, 3 months ago)

Nice SAB,  you would laugh if you saw mine!


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Offlinesocraticd
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: liamtheloser]
    #19079688 - 11/03/13 10:41 AM (10 years, 3 months ago)

Haha.  Thanks guys.  I have a lot to learn but I figure with equipment, anything worth doing is worth doing right.  The SAB is honestly pretty cheap and easy, and only took an hour or so to put together.  Definitely worth it.


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OfflineForresterM
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: socraticd]
    #19079797 - 11/03/13 11:02 AM (10 years, 3 months ago)

I should do one right some time, it would probably help my contam rate...
Mine's just half circles cut at the top of a tub then I flip it upside down on top of a damp towel for working.


--------------------
Repugnant is a creature who would squander the ability to lift an eye to heaven, conscious of his fleeting time here.
-------------------

Have some medicinal mushrooms and want to get the most out of them?  Try this double extraction method.


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Invisibleliamtheloser
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: Forrester]
    #19080083 - 11/03/13 12:17 PM (10 years, 3 months ago)

Since I can't stand to waste a tote by cutting holes in it, i just flip a tote on its side.  It's open faced but I rarely get a contaminated plate, in fact, out of the last 100 or so, I haven't gotten a contamination (except from infected wedges I got from someone).


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Offlinesocraticd
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: liamtheloser]
    #19084860 - 11/04/13 10:39 AM (10 years, 3 months ago)

A gfresh sleeve of plates poured last night, de-condensing in the SAB for a few days before my new edible cultures show up.



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Offlinesocraticd
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: socraticd]
    #19091847 - 11/05/13 07:06 PM (10 years, 3 months ago)

Did a bit more agar work tonight.  I received a trio of syringes from The Spore Depot:

- King Oyster
- Pearl Oyster
- Enoki

I transferred one drop from each syringe on to 3 plates to start growing those out.

Additionally, I made my first round of potential master slants out of two possible vessels.  I purchased some of these 20mmx150mm culture tubes, and didn't realize the necks on them would be as narrow as they were.  Not nearly wide enough for a scalpel.  I also purchased an inoculation loop, so I figured I'd give them a shot anyway.  I also purchased a case of the wider amber vials you see in this pic, which are plenty wide enough for a scalpel blade.







Ultimately, here are the results of the night's work.  Three plates each of Enoki, Pearl Oyster, King Oyster, Yellow Oyster.  Three attempts at a start of a P3 master slant of Reishi in the 20x150mm tubes, and four attempts at a master slant of Reishi in the wider amber vials.



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OfflineJeff
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: socraticd]
    #19091909 - 11/05/13 07:16 PM (10 years, 3 months ago)

I have never had an issue with scalpels fitting into a 20 X 50 but I do fill them a lot more than what is shown in your pic.  Probably close to 3X more.  I use a #11 blade and it is narrow enough to manipulate in a 20X.
Either way, for both pictured, I would use more agar.  For the 20 X 150's I like to have the agar, once slanted, to be about at the 50% mark on the lower end and about 30% on the amber vial.


--------------------
Myco-tek


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Offlinesocraticd
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Re: Socraticd's experiments - a.k.a, a newbie's grow log [Re: Jeff]
    #19092515 - 11/05/13 09:03 PM (10 years, 3 months ago)

Hmm, I literally can't imagine a scalpel blade of any size fitting in to these tubes.  I'll snap a pic tomorrow morning of one of the empty ones next to one of my exacto's for comparison.

The amber ones seem easy to work with, though, and i bought a whole case of them. 

One question I did have, though, is about condensation in the slant tubes.  Unless I devise a way to PC them at the proper angle to begin with, I have to PC them then remove once cool enough to handle, and let them finish cooling outside the PC, which will leave the pictured amount of condensation. 

Suggestions on a way around this, and/or should I even worry?  I mean, my longer term experimentation will probably give me the answer, but suggestions/anecdotes are always welcome.


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