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InvisibleSpunkyMonkey88
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Registered: 10/08/19
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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26399138 - 12/24/19 01:12 PM (4 years, 1 month ago)

Quote:

Shr00merN00ber said:
Just got done pouring and wrapping with parafilm. Man that parafilm can get annoying with cutting and stretching etc. Here is how it looks. I dont know why I got alot of condensation on some plates (not all), I had the bottle in a water bath at 47C for a while before. Some dishes have these bits in it, all good or  ones with bits looking trash? I seem to find the pieces settled on the bottom.









Cheers!




Looks good man! And dont worry about the sediment itll still function just the same...

I'm weird about that too so if you want to avoid that next time make twice as much LME as you need and put in a canning jar in the fridge over night, then siphon the top half off with what ever you can (i used a Turkey baster) then run that through a coffee filter, then add your agar for some clear ass agar, If that's what you're looking for...

Anyway that whole process is purely for astectic reasons and you could probably skip either the coffee filter or the s
Mason jar/ siphon step and still get pretty clear agar...

Good work man pouring plates is a huge milestone!


Edited by SpunkyMonkey88 (12/24/19 01:16 PM)


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InvisibleSpunkyMonkey88
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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26399142 - 12/24/19 01:14 PM (4 years, 1 month ago)

Also the coffee cup with hot water ontop of your plates trick works like a charm for the condensation :wink:


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OfflineShr00merN00ber
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Registered: 12/19/18
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Re: First time will attempt cloning [Re: SpunkyMonkey88]
    #26404095 - 12/28/19 05:17 AM (4 years, 1 month ago)

Hey guys, hope you are doing well. So, I've just noticed this on one of my plates. It seems like a very small side colony, maybe its contam or a tiny piece of mycelium that fell off during the agar transfer. :crazy:



So my question is, if I want to check the petris in greater detail, do I need to go back to the SAB and take them out of the ziplock bags in there? All dishes are wrapped in parafilm. I dont want to open them, just check each individual dish in detail with a light shining on them. Right now I'm just trying to see them through the outside of the bags, lifting each one and checking.

Thanks!


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OfflinePhycus
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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26404238 - 12/28/19 08:12 AM (4 years, 1 month ago)

if everything is wrapped in parafilm, you really shouldnt need the ziplocs.


--------------------
disclaimer - nothing i post is real. this account is for fictitious purposes and posts should not be taken literally


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Offlinetrubblesome
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Re: First time will attempt cloning [Re: Phycus]
    #26404253 - 12/28/19 08:23 AM (4 years, 1 month ago)

yeah you could play frisbee with one if you wanted as long as it's got something keeping it closed.

you don't have to wrap them until you've put something on it, otherwise you're going to waste a lot of parafilm - once they're set stack them and put the sleeve they came in over the stack. flip them over, fold a paper towel in to a square and put it on top, then tape the sleeve shut. the paper towel will help cut down on condensation. You can stack them upside down before putting the sleeve over the stack so that they're right side up in the bag, but I like taking them out of the bag better when they're in it upside down as it's more difficult to accidentally remove the top of one and potentially contaminate it.


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: trubblesome]
    #26404358 - 12/28/19 09:39 AM (4 years, 1 month ago)

Here are some pics from two of my petris. To be honest, I didn't wash my hands and handled them 'carefully' with my bare hands. Here is a plate I feel like could be contaminated to my untrained eyes.

Both the following and the previous plate I posted came from the same cluster tissue sample:

Front



Back



Back zoom




Some info from experienced eyes would be nice.



Here is another petri just for fun from the big clone tissue sample.

Front



Back



Cheers!


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OfflineShr00merN00ber
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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26405106 - 12/28/19 05:34 PM (4 years, 1 month ago)

Got a couple of questions I'm wondering about (sorry for the double post). What is the best method for spawning bulk while trying to isolate genetics.. g2g or lc?

What is the general consensus on the best method when taking speed and reliability (less chance of contam) into consideration?


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InvisiblefahtsterM
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Re: First time will attempt cloning [Re: Shr00merN00ber]
    #26405284 - 12/28/19 07:23 PM (4 years, 1 month ago)

G2g will be a bit faster but it’s riskier because you’re opening the jar/bag... LC can be inoculated through a self-healing port. 

Agar to agar transfers are your best bet for isolation.. not sure how lc’s and g2gs are going to be better than one another for isolating.. if you mean which is better for reducing genetics from an MS inoculation, I’d say g2gs since they’re somewhat stationary whereas, lcs are free floating in liquid; you’ll end up with more sub strain colonies.

Since you already have agar going, look into https://www.shroomery.org/forums/showflat.php/Number/24740168/fpart/1/vc/1 josex biopsy method for LCs (sorry if it’s already been said—I’m on the move)

Faht


Edited by fahtster (12/28/19 07:33 PM)


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OfflineShr00merN00ber
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Registered: 12/19/18
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Re: First time will attempt cloning [Re: fahtster]
    #26405614 - 12/29/19 02:27 AM (4 years, 1 month ago)

Awesome, thanks for the info Faht. Do you see any problems with the petris above? In a couple days I'll post a collage with all petris so maybe someone can help me choose the best plate for g2g.


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