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 tryptkaloids
Learner
Registered: 02/08/15
Posts: 12,654
Loc: Exact Center
Last seen: 1 day, 11 hours
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 Re: Pasty Agar Tek  [Re: insutama]
#25375308 - 08/08/18 03:32 PM (5 years, 8 months ago) |
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Ive left them on for 3 years now with 0 issue, paper its definately due to small holes.
Quote:
insutama said:
i took the paper towel off im thinking mine arnt drying up because i made 1/8" hole for FAE instead of the 1/4" it said in the tek and i put 3 layers of micropore tape on it. Im getting some real healthy looking mycelium on my agar just so much water pooling up now
To aid pooling use them upside down, pooling is bad
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TaderHoagie
Hypernormalized
 
Registered: 10/21/17
Posts: 229
Last seen: 5 years, 4 months
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I was able to do this from memory! I'm happy about that.
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Ginge
Sponge Brain
Registered: 07/08/18
Posts: 202
Loc: Rocky Mountains
Last seen: 5 years, 2 months
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So I wanted to be more fancy and ordered some petrie dishes. When using pasty plates I lost 1/36 due to contams when nocing with MS syringes. And over the course of 60 more plates lost 2 others to contamination. Pretty good, I thought. So I am trying to start PE in the petries. I see contams and myc on most of the plates, 10/16. I used the technique of PCing my agar in a jar, then pouring the plates in two stacks of ten, starting from the bottom in my SAB. Left them in the SAB overnight and did the MS work in the morning. Used a homemade Bod's inoculation loop flamed before every plate. The contamination is so great I want to blame the syringe, but I think that may be vein as my success sample is pretty small. I was thinking of cooking up some pasty plates and nocing them with the syringe to try and test the theory. And after typing this as a question I feel like that would be a pretty good test, sometimes it helps just to type it out, but I will still post, maybe somebody has a similar thought.
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Ginge
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verum subsequentis
seeker of truth
Registered: 03/22/16
Posts: 8,732
Last seen: 1 year, 10 months
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Re: Pasty Agar Tek [Re: Ginge]
#25390976 - 08/15/18 07:33 PM (5 years, 7 months ago) |
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if you are wondering about your pouring tek then just wrap a few plates directly after pouring and see if they are still clean a week later. if not you are fudging up on your pouring if so, you either contammed during the noc up or your syringe is problematic
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pixelpopper
Crap Artist
Registered: 09/20/13
Posts: 4,022
Loc: Dreamland
Last seen: 5 months, 27 days
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Re: Pasty Agar Tek [Re: Ginge]
#25390978 - 08/15/18 07:34 PM (5 years, 7 months ago) |
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If you're losing most of your plates to contams but not getting the same rate of contams on pasty plates, using the same syringe, then something is probably off with your plate technique
If you're using different syringes, then comparison can't really be made.
In any case, you can still transfer healthy myc away from the contam plates to fresh plates
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 flyhighfunguy
Registered: 09/13/17
Posts: 1,550
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If I pressure cook a batch of pasty plates in the morning do you guys think they'd be cool enough to inoculate by night time?
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foragedfungus
Registered: 09/30/13
Posts: 1,867
Loc: out there
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There's not much thermal mass to a pastey plate. They're cool enough to use less than an hour after they come out of the cooker. Pretty much as soon as the agar solidifies.
Edited by foragedfungus (08/17/18 06:06 PM)
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Ginge
Sponge Brain
Registered: 07/08/18
Posts: 202
Loc: Rocky Mountains
Last seen: 5 years, 2 months
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Yeah Verum, thank you. I will definitely test my pouring technique this weekend with a new batch of agar. I will fridge the other plates for ten days while I observe the test plate. *edit* This morning I made 20 plates, kept the top two with the most condensation as my tests, and noc'd up the remaining 18. I also had 4 pasty plates which I also noc'd. (both with the same MS syringe as before) In hindsight it was foolish and cocky to go to petries without running a test on my pour technique
I realize I should have said that I would be using the same syringe for the test. Such common sense to me but, well yeah, I will be more specific next time.
*edit* I almost always review TEKs before I work. In this case, with a massive contamination problem, I reviewed SAB technique, agar making and agar pouring technique. Upon review I made 1 mistake I knew about, and several that I didn't.
1-I had a towel and no rack for my SAB bottom, the petries were way to close to the towel.
2-I was only flaming my loop after every plate during the transfer because I wasn't touching the syringe to the loop, stupid!, I should have been flaming both every time.
3-I was not streaking the plate correctly. This didn't contribute to contams, but might have created more situations with viable myc that could be transferred.
4-The mistake I knew about, not realizing how significant it was. I was trying to pour from the bottom of the stack up. At one point I lifted the base of the next plate, and didn't see it through the condensation, and poured onto a lid. I knew I messed up but didn't think about how that created a vector for every lower plate to essentially touch the towel. I should have called this a learning experience at that point but I tried to use the good plates from the pour.
Kinda of embarassing but hopefully someone learns from it and avoids the same mistake.
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Ginge
Edited by Ginge (08/18/18 08:08 PM)
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Johnny.rotten
Traveller
Registered: 08/16/18
Posts: 159
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Re: Pasty Agar Tek [Re: Ginge]
#25398804 - 08/19/18 03:15 PM (5 years, 7 months ago) |
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Thank you for the great write-up. Without it, I wouldn't of gotten into agar. I have a round of transfers to do this evening. Then i'm preparing another run of plates for the next round.
  
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Shr00mMan
Stranger
Registered: 07/11/18
Posts: 10
Last seen: 2 years, 1 month
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As a noon I love this tek... THANKS FOR THE INFO FOSHO
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drprobablyno
Good Person
Registered: 08/22/18
Posts: 76
Last seen: 3 years, 11 months
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Thanks. I will make this a part of my guide 
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drprobablyno
Good Person
Registered: 08/22/18
Posts: 76
Last seen: 3 years, 11 months
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Re: Pastywhyte's Easy Agar Tek [Re: Pastywhyte]
#25404860 - 08/22/18 09:39 AM (5 years, 7 months ago) |
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Thanks Pasty....Very useful. Will be part of my guide
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DcevLearn
Team Tomentose
Registered: 07/07/18
Posts: 330
Last seen: 7 months, 16 days
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On the topic of condensation... I caught up on the last 30 pages trying to solve my issues with condensation. It seemed like most of the recommendations were waiting for the agar to cool in the dishes before pressure cooking and to use glass jars.
I tried the following:
- Glad Mini Rounds
- Ball Short Half Pint Jelly Jars w/ micro pore vent
- Ball Dry Herb Jars w/ plastic lid unscrewed slightly
- Used leftovers as pour agar to petris in SAB
The mini rounds and the petris ended up with considerable condensation but the glass options both had barely any.
My personal favorite setup is the dry herb jars and plastic storage caps. They don't have any weird edges so they are easy to clean. They have less distortion than the jelly jars, but be aware there is still distortion, these aren't crystal clear petris.
Has anyone added vents to the plastic storage lids and do you have any feedback if that helped with clearing condensation over time?
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Current Project: LAGM: Stones From Scratch in 2020
   
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LtLurker
Lost Sailor
Registered: 01/03/18
Posts: 7,535
Loc: Borderlands 
Last seen: 1 month, 13 hours
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Re: Pastywhyte's Easy Agar Tek [Re: DcevLearn]
#25412377 - 08/25/18 11:59 AM (5 years, 7 months ago) |
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Neat-o. I wonder why the glass is clear and plastic isn't? The pour agar plates should be clear as long as it's not poured too hot.
I can't fully solve the condensation problem with my pasty's, but what I've been doing is letting my twist-n-locks cool in the pc so they have a bit of a slant, then store upside down. When I'm ready to use it, most condensation has settled on the lid, open upside-down, turn lid over to empty out pooled water, do my work. This is getting most of my excess moisture out. Any moisture left behind will pool at the lowest end due to the agar's slant and doesn't run across the plate.
I haven't tried adding vents to them. Unmodded screw on lids can be backed off an 1/8th a turn to allow some exhaust while pc'ing, then tightened at the end of the cycle.
Edited by LtLurker (08/25/18 12:03 PM)
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flyhighfunguy
Registered: 09/13/17
Posts: 1,550
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Re: Pastywhyte's Easy Agar Tek [Re: LtLurker]
#25419452 - 08/28/18 03:01 PM (5 years, 7 months ago) |
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Do you guys ever have trouble getting a transfer off of your blade? I transfered some plates a couple nights ago, and noticed that some of the transfers were getting stuck on the blade. It would sometimes be completely stuck on the blade, so i would just re-flame the scalpel and go for another transfer, and other times i would try to wiggle it off the blade and it would just end up falling on the floor of my SAB.
Any recommendations on how to fix this? I use 2% Telephone agar, 2% LME, and each plate usually gets between 1.5 - 1.75 teaspoons of agar.
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LtLurker
Lost Sailor
Registered: 01/03/18
Posts: 7,535
Loc: Borderlands
Last seen: 1 month, 13 hours
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Usually just moving my blade the opposite direction I went to pick up the wedge makes it slide right back off. Keep it to the tip of your scalpel as much as possible.
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JHOVA
Post whore
Registered: 02/17/17
Posts: 4,727
Loc:
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Re: Pastywhyte's Easy Agar Tek [Re: LtLurker]
#25419641 - 08/28/18 04:17 PM (5 years, 7 months ago) |
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Practice in open air. Its easier to do a transfer if the wedge is closer to the tip. Try to cut the recipient agar plate just barely slicing the surface. Thats what works for me. When i cut deep into recipient agar the wedge gets stuck higher up the blade.
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Munchauzen
Registered: 06/22/11
Posts: 14,344
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Quote:
flyhighfunguy said:
Do you guys ever have trouble getting a transfer off of your blade? I transfered some plates a couple nights ago, and noticed that some of the transfers were getting stuck on the blade. It would sometimes be completely stuck on the blade, so i would just re-flame the scalpel and go for another transfer, and other times i would try to wiggle it off the blade and it would just end up falling on the floor of my SAB.
Any recommendations on how to fix this? I use 2% Telephone agar, 2% LME, and each plate usually gets between 1.5 - 1.75 teaspoons of agar.
ya i try to transfer the wedges with just the very tip of the blade, so it takes very little effort to make it fall off
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stareatclouds
star eat clouds?
Registered: 09/29/14
Posts: 10,126
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Re: Pastywhyte's Easy Agar Tek [Re: Munchauzen]
#25420187 - 08/28/18 08:19 PM (5 years, 7 months ago) |
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In addition to what Munch and JHOVA said above, it helps me to have a less sharp blade (one I've used a bit). Another thing I'll do when the plates are thicker is slice horizontally under the myc after I've cut a triangle/square outline. This rests it sideways/flat on the top of the blade, making it easier to come off in the receiving plate.
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bakenast
Muffen Stuffer
Registered: 03/14/11
Posts: 920
Last seen: 1 year, 3 months
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Re: Pastywhyte's Easy Agar Tek [Re: Pastywhyte]
#25431449 - 09/02/18 07:50 PM (5 years, 7 months ago) |
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I posted this question in another thread but this thread looks like where it originated , anyways “ Has anyone every used a syringe filters on no pour plates instead of micropore. I have 100s of them but no micro pore atm wondering if they would work instead with high temp silicone holding them..
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