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Turvenuija
Up shroom creek without a paddle

Registered: 11/07/18
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First attempt at agar is a bit... off
#26612109 - 04/19/20 05:54 AM (3 years, 9 months ago) |
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So I made some plates with 20g/l brf and agar mix, Bod's agar tek warned that brf can have some sediment issues but is this clumping acceptable? I also messed up with my SAB, I used the tub lid as the floor and that turned out to be shit for pouring agar because it wasn't flat enough. I poured when the media was still too hot so I got a lot of condensation. I didn't see what was happening before the plates had already solidified so I'm stuck with these weird plates.
I inoculated three of these with spores using a diy loop, I hope I can at least get a transfer out of them.
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PiggyPig
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Re: First attempt at agar is a bit... off [Re: Turvenuija]
#26612140 - 04/19/20 06:33 AM (3 years, 9 months ago) |
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If your agar is too hot when you pour it, it will cause more condensation inside the dish. Once they are poured, leave them stacked and place a cup of hot water on top of the stack. This will also help.
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redhandmat
Dude


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Re: First attempt at agar is a bit... off [Re: Turvenuija]
#26612150 - 04/19/20 06:40 AM (3 years, 9 months ago) |
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Quote:
Turvenuija said: So I made some plates with 20g/l brf and agar mix, Bod's agar tek warned that brf can have some sediment issues but is this clumping acceptable? I also messed up with my SAB, I used the tub lid as the floor and that turned out to be shit for pouring agar because it wasn't flat enough. I poured when the media was still too hot so I got a lot of condensation. I didn't see what was happening before the plates had already solidified so I'm stuck with these weird plates.
I inoculated three of these with spores using a diy loop, I hope I can at least get a transfer out of them.

I think we almost always botch our first attempts. So dont worry too much that it didnt become perfect. I dont think the clumping will be a big of an issue. What is more important is that you had a good sterile tek and didnt move air too much in your SAB, never hovering your hands over the dish etc. Of course PCing for the appropriate time length and 15 psi.
Condensation is a bitch, but what helps is doing what piggypig told you above. After that the only thing you can really do is store the dish upside down and let condensation pool on the glass/plastic/lid instead of on the agar.
Edited by redhandmat (04/19/20 06:42 AM)
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Sockadin



Registered: 01/03/10
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Re: First attempt at agar is a bit... off [Re: redhandmat]
#26612161 - 04/19/20 06:49 AM (3 years, 9 months ago) |
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Condensation is part of the process. I see so many people try to come up with ways to minimize it,but clean transfers and storage are way more important than getting to condensation out. I wouldn't flip them or put a hot cup of water. Good gas exchange will allow the condensation to evaporate in a couple of days.
Also I like to use a cookie drying rack to elevate my plates and media off of the surface of the SaB. Helps tremendously.
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Turvenuija
Up shroom creek without a paddle

Registered: 11/07/18
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Re: First attempt at agar is a bit... off [Re: Sockadin]
#26612180 - 04/19/20 07:03 AM (3 years, 9 months ago) |
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I filled a 2 liter zip lock bag with hot water and covered my dishes with it, the only reason is to be able to see what's happening.
Quote:
Sockadin said: Good gas exchange will allow the condensation to evaporate in a couple of days.
How can plates have gas exchange when they're closed and sealed with cling wrap?
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redhandmat
Dude


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Re: First attempt at agar is a bit... off [Re: Sockadin]
#26612185 - 04/19/20 07:05 AM (3 years, 9 months ago) |
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Quote:
Sockadin said: Condensation is part of the process. I see so many people try to come up with ways to minimize it,but clean transfers and storage are way more important than getting to condensation out. I wouldn't flip them or put a hot cup of water. Good gas exchange will allow the condensation to evaporate in a couple of days.
Also I like to use a cookie drying rack to elevate my plates and media off of the surface of the SaB. Helps tremendously.
Im sure you are more experienced, but proper storage IS to have them upside down. At least according to people like RR etc.
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LadysKnight
Hello Ladies


Registered: 10/09/15
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Re: First attempt at agar is a bit... off [Re: Sockadin]
#26612186 - 04/19/20 07:05 AM (3 years, 9 months ago) |
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Upside down is the proper orientation for a SAB. To fix the uneven lid issue, remove it and place the SAB on a hard flat surface like a table.
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Turvenuija
Up shroom creek without a paddle

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Re: First attempt at agar is a bit... off [Re: LadysKnight]
#26612199 - 04/19/20 07:13 AM (3 years, 9 months ago) |
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Yeah using the lid as a floor is fine for doing pf tek but for future agar work I'll have to ditch the lid.
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Sockadin



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Re: First attempt at agar is a bit... off [Re: Turvenuija]
#26612232 - 04/19/20 07:28 AM (3 years, 9 months ago) |
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Flip the SAB not the plates.
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PinkStormtrooper
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Re: First attempt at agar is a bit... off [Re: Sockadin]
#26612237 - 04/19/20 07:34 AM (3 years, 9 months ago) |
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that hot water cup trick didnt work for me. ended up waiting a day then opening the plates upside down inside the SAB and using an iso wipe to collect the water droplets then close the plates back up.
-------------------- "say, you got a little astroglide on your moustache"
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redhandmat
Dude


Registered: 05/09/19
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Quote:
PinkStormtrooper said:

that hot water cup trick didnt work for me. ended up waiting a day then opening the plates upside down inside the SAB and using an iso wipe to collect the water droplets then close the plates back up.
Thats balsy. You might have contaminated them by wiping inside that way. letting water collect on lid (instead of on the agar) and leaving it to evaporate away with time wouldnt hurt anything.
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Htaeh
Immortal


Registered: 10/14/18
Posts: 320
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Lol hope they're okay! Someone suggested putting the plates upside down on their laptop for ten minutes normally clears it up for them.
-------------------- "To write is to wage war" ~ Voltaire "The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D. "A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher "We'll know our disinformation program is complete when everything the US public believes is false." ~ William J. Casey, 1981 "Fear always works to influence the population, when the population is uneducated." "The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~ Maximilien Robespierre "Knowledge makes a man unfit to be a slave." ~ Frederick Douglass
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poisoned
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Re: First attempt at agar is a bit... off [Re: Htaeh] 1
#26612746 - 04/19/20 11:24 AM (3 years, 9 months ago) |
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Don't do that in the future. Iso is not sterile, neither are any wipes dipped in it.
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Machiavelliavore
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Re: First attempt at agar is a bit... off [Re: poisoned] 1
#26612771 - 04/19/20 11:35 AM (3 years, 9 months ago) |
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If there are a lot of clumps you have have effectively reduced the nutritional content of your agar to some degree, but at 2% brf that will be no issue. Also, I personally find I get less clumping when I add hot water to my agar jar first, then whisk in the dry ingredients, as opposed to lots of clumping when putting dry ingredients and putting agar on top.
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I spawned some popcorn casings and had double-overlay cause I didn't put enough hydrogen peroxide in my automated aquarium mister. I only got one mushroom so I cut off the head part where the seeds fall from and put it in a jar of LC and sprayed it all over a tin of PF cakes I made with gravel, cardboard, and bisquick in my microwave. I think it will be good cause B+ is so potent. Triggered yet? Only a square would say "a cube is a cube."
No, this does not look right...
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mushhead
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Re: First attempt at agar is a bit... off [Re: redhandmat] 1
#26612792 - 04/19/20 11:41 AM (3 years, 9 months ago) |
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Quote:
redhandmat said:
Quote:
PinkStormtrooper said:

that hot water cup trick didnt work for me. ended up waiting a day then opening the plates upside down inside the SAB and using an iso wipe to collect the water droplets then close the plates back up.
Thats balsy. You might have contaminated them by wiping inside that way. letting water collect on lid (instead of on the agar) and leaving it to evaporate away with time wouldnt hurt anything.
I have done this and I will confirm that I did have contamination in my plates, one bacterial and a really bad mold issue after iso wiping the lids. Its easier just to allow it to evaporate, I have left plates upside down with no real change unless left on a heating apparatus of some kind - Here's why that works in case anyone was wondering: You're just running physics in reverse and allowing the condensation to re-condense on the agar itself. Is this more clean than just allowing it to evaporate? discuss.
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Meditation Principles Silence: Giving you room to listen / Stillness: Giving you room to feel / Spaciousness: Just giving you room
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redhandmat
Dude


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Re: First attempt at agar is a bit... off [Re: mushhead]
#26612860 - 04/19/20 12:07 PM (3 years, 9 months ago) |
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Quote:
mushhead said:
Quote:
redhandmat said:
Quote:
PinkStormtrooper said:

that hot water cup trick didnt work for me. ended up waiting a day then opening the plates upside down inside the SAB and using an iso wipe to collect the water droplets then close the plates back up.
Thats balsy. You might have contaminated them by wiping inside that way. letting water collect on lid (instead of on the agar) and leaving it to evaporate away with time wouldnt hurt anything.
I have done this and I will confirm that I did have contamination in my plates, one bacterial and a really bad mold issue after iso wiping the lids. Its easier just to allow it to evaporate, I have left plates upside down with no real change unless left on a heating apparatus of some kind - Here's why that works in case anyone was wondering: You're just running physics in reverse and allowing the condensation to re-condense on the agar itself. Is this more clean than just allowing it to evaporate? discuss.
Yes makes sense that would ahppen. My personal opinion is that water doesnt hurt anything if its not on agar.... so why even bother with it? just store the petri dish upside-down (as it should be in any case) and it will go away by itself.
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Htaeh
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Re: First attempt at agar is a bit... off [Re: redhandmat]
#26612869 - 04/19/20 12:11 PM (3 years, 9 months ago) |
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*quickly runs to the area plates are stored to flip them all*
-------------------- "To write is to wage war" ~ Voltaire "The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D. "A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher "We'll know our disinformation program is complete when everything the US public believes is false." ~ William J. Casey, 1981 "Fear always works to influence the population, when the population is uneducated." "The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~ Maximilien Robespierre "Knowledge makes a man unfit to be a slave." ~ Frederick Douglass
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redhandmat
Dude


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Re: First attempt at agar is a bit... off [Re: Htaeh]
#26612878 - 04/19/20 12:15 PM (3 years, 9 months ago) |
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Quote:
Htaeh said: *quickly runs to the area plates are stored to flip them all*

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orange771
Registered: 01/26/11
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Re: First attempt at agar is a bit... off [Re: Htaeh]
#26612884 - 04/19/20 12:16 PM (3 years, 9 months ago) |
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I just make plates and let them sit in the SAB unwrapped for a few days until there’s almost no condensation. Never been much of a problem.
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LadysKnight
Hello Ladies


Registered: 10/09/15
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Re: First attempt at agar is a bit... off [Re: orange771] 1
#26612900 - 04/19/20 12:21 PM (3 years, 9 months ago) |
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Quote:
RogerRabbit said:
No. You should not store petri dishes upside down. It forces the water in the agar to 'drain' out, collecting on the lid, which then floods the culture when you right the dish/jar. In addition, mycelium grows best on the top(away from gravity) surface of a flat plane, and when forced to grow upside down, growth and performance is slower. Since the whole point of isolation on agar is to encourage the fastest growth possible, it should be done in actual Petri dishes that are stored right-side-up for best results. RR
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