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Dactylium
Don't touch me I'm sterile


Registered: 08/12/16
Posts: 696
Last seen: 10 months, 13 days
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Re: When to give up on Agar Plates??? [Re: bodhisatta]
#24022625 - 01/19/17 10:48 AM (7 years, 11 days ago) |
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Yeah you can get a pocket scale for small stuff like this for $6.50 on amazon
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Mushenstein
Brain Damage



Registered: 02/19/15
Posts: 680
Loc: At The Gates
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Re: When to give up on Agar Plates??? [Re: RealityCzech]
#24022721 - 01/19/17 11:32 AM (7 years, 11 days ago) |
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Thanks for the directions. I had no idea they were so cheap. Back to my last question, if anyone knows.
Quote:
RealityCzech said:I aspirate off the extra uncolonized lc, then pull out the mycelium and mix it with about 50 ml of distilled water.
How do you aspirate excessive LC off of the Myc in an LC container? Is this even possible or is this guy full of it???
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Mycolorado
Hobbyist


Registered: 07/23/16
Posts: 8,529
Loc: Interdimensional Bootcamp
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Re: When to give up on Agar Plates??? [Re: Mushenstein]
#24022728 - 01/19/17 11:37 AM (7 years, 11 days ago) |
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Quote:
Mushenstein said: Thanks for the directions. I had no idea they were so cheap. Back to my last question, if anyone knows.
Quote:
RealityCzech said:I aspirate off the extra uncolonized lc, then pull out the mycelium and mix it with about 50 ml of distilled water.
How do you aspirate excessive LC off of the Myc in an LC container? Is this even possible or is this guy full of it???
Yes, this dude is clueless.
Just eyeball it until you get a proper scale. I'd also just let those plates ride a while as suggested...they might pop eventually....
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RealityCzech
Entropy enthusiast


Registered: 12/29/16
Posts: 90
Last seen: 6 years, 3 months
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Re: When to give up on Agar Plates??? [Re: Mushenstein]
#24022934 - 01/19/17 01:26 PM (7 years, 11 days ago) |
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Quote:
Mushenstein said: How do you aspirate excessive LC off of the Myc in an LC container? Is this even possible or is this guy full of it???
The myc tends to stay in a little cloud in the LC near the bottom. There is a lot of "clear" media that remains uncolonized. Use a large syringe through your port to suck the clear media off the top. Once you get as much as you can without getting the myc cloud, use a second syringe to draw out the mycelium and stop when you've got most of it leaving some of the "clear" media that was in the bottom behind.
I'll then add the myc to distilled water in a small bottle with a SHIP. Right now I'm using some cobalt blue dropper bottles that I flipped the dropper bulb over and filled the bulb with silicon. I don't know if it is necessary, but I PC the distilled water in another bottle and PC the bottle with 10 ml in it. Once cooled, I close them up so there isn't a vacuum and add the myc and distilled water that was PCed separately.
It doesn't get ALL the liquid media out, but with the distilled water it dilutes it quite a bit. Once well shaken it will break up the mycelium and allow you to easily draw and inject it. It isn't super thick like some of the LC syringes that you can buy, but there is plenty of myc and it keeps for a long time in the fridge.
Use the bottle like you would a med draw... if you're adding liquid to it, draw out 1 ml of air from the top for every ml you injected after injection. When you go to draw liquid out, fill your syringe with air and inject it before you draw out the liquid (again, equal volume of air going in and liquid coming out). This will keep the pressure in the bottle relatively stable over time... At least long enough to use up the LC.
I am not home at the moment, but I've got a jar of golden oyster LC that's just about ready for this at home. I'll snap a pic of that and of one of the cobalt bottles that's running low tonight and post them so you can see what I mean.
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RealityCzech
Entropy enthusiast


Registered: 12/29/16
Posts: 90
Last seen: 6 years, 3 months
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Re: When to give up on Agar Plates??? [Re: RealityCzech]
#24023162 - 01/19/17 02:58 PM (7 years, 11 days ago) |
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As promised... Note how the mycelium is sitting on the bottom of the jar? Use that to your advantage and use a syringe to aspirate off the extra liquid before you pull the myc out.

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Mushenstein
Brain Damage



Registered: 02/19/15
Posts: 680
Loc: At The Gates
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Re: When to give up on Agar Plates??? [Re: RealityCzech]
#24023163 - 01/19/17 02:59 PM (7 years, 11 days ago) |
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I was under the impression that an LC isn't done until the water is 90% filled with the myc. Your saying that this is having too high of an expectation??? When do you consider the LC done and ready to go to syringe?
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RealityCzech
Entropy enthusiast


Registered: 12/29/16
Posts: 90
Last seen: 6 years, 3 months
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Re: When to give up on Agar Plates??? [Re: Mushenstein]
#24023208 - 01/19/17 03:14 PM (7 years, 11 days ago) |
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I can't speak to other media recipes, but with the turbinado spores seem to germinate in 2-3 days and it slows to a crawl about 5 days later with 33-50% of it colonized. Maybe I'm impatient and could get more, but when I don't see noticeable growth for a couple days I'll go ahead and pull it, run it on agar for contaminates, and call it a day.
There are a lot of little variables and your results may vary... I'm just giving you the easiest thing I've found. If it helps, great. If not, that's fine too.
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Mushenstein
Brain Damage



Registered: 02/19/15
Posts: 680
Loc: At The Gates
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Re: When to give up on Agar Plates??? [Re: RealityCzech]
#24023811 - 01/19/17 07:00 PM (7 years, 11 days ago) |
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When should I transfer from these plates? Do I wait until they are 100% colonized or does it matter. They will be totally done in a few days, but if it doesn't matter I will cut um up now!
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Voltageloss
Stranger everyday

Registered: 11/23/16
Posts: 79
Loc:
Last seen: 5 years, 3 months
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Re: When to give up on Agar Plates??? [Re: Mushenstein]
#24024161 - 01/19/17 08:51 PM (7 years, 11 days ago) |
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New to agar but it's my understanding you want to transfer from a leading edge, so full colonization is not the goal.
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Stop shopping at Walmart
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synthetik
biological entity

Registered: 12/15/16
Posts: 194
Last seen: 3 years, 10 months
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Re: When to give up on Agar Plates??? [Re: Voltageloss]
#24024581 - 01/20/17 12:37 AM (7 years, 10 days ago) |
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^this is my understanding as well, Ive also heard this is because it touching the sides can be a contam vector... not sure how esp in a pasty plate that should be 100% sterile, but as far as ive seen people tiger drop at about 80% colonization
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Mushenstein
Brain Damage



Registered: 02/19/15
Posts: 680
Loc: At The Gates
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Re: When to give up on Agar Plates??? [Re: Voltageloss]
#24024981 - 01/20/17 08:28 AM (7 years, 10 days ago) |
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Quote:
Voltageloss said: New to agar but it's my understanding you want to transfer from a leading edge, so full colonization is not the goal.
This is sound logic. Unless someone argues differently I will cut from the leading edge. Since these are transfers, the plates that colonize from these cuts will be going to grain, so avoiding contam factors is paramount. I'd rather not have to clean it up again.
I have become very impressed with the performance of Dog Food Agar as opposed to Agar recipes with more elegant ingredients. I made some more last night and had a slight accident with the addition of food coloring, in which much more was added then should have been. like 20 drops as opposed to 3. I wonder if it will have any impact on sustaining the growth or germination of fungal strain.
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themadextractor
Stranger


Registered: 06/25/14
Posts: 230
Last seen: 1 year, 3 months
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Re: When to give up on Agar Plates??? [Re: Mushenstein]
#24025737 - 01/20/17 02:01 PM (7 years, 10 days ago) |
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Definitely transfer now. That looks nice and clean. You do want the leading edge. It's the freshest mycelium.
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