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tijuanataximan
Stranger

Registered: 07/09/20
Posts: 32
Loc: cananananada
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Issues with my hot pour agar technique
#27031207 - 11/10/20 08:14 AM (3 years, 3 months ago) |
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There are a number of problems that I am having with my hot pours. The major ones include:
- Condensation
I put my media bottle in a water bath with a thermometer on the side. I begin pouring at approximately 120f or 48c and 75% of my plates (15/20) still have excess condensation on the top and sides.
- Lack of growth/stunted growth
I have read through and followed BOD's Comprehensive Agar TEK as well as a few variations of the more popular agar recipes (Mainly PDA/Y or subbing in corn starch in place of potato starch and with varying amounts of additives but staying within the guidelines posted here, in Psilicon's write up and yet almost every plate I pour ends up with no growth or very stunted growth. I even had a plate show trich that didn't get any bigger than 2mm LOL
 Note the discoloration around the area that I had cut out. Looking closely it seemed as though very small mycelial strands were running through the area and sucking up the food coloring
 I believe this was from the type of yeast I had attempted to use to see if it made a difference. It didn't.
I am now preparing three new media bottles to try which include a grain soak water, one with potato starch using distilled water, and one with corn starch using distilled water (I normally use my local tap water). At this point I'm starting to think that the issue could very well be with the sample(s) that I am working with which include 4 spore syringes purchased from a vendor and a clone from my very first grow (pre-made monotub from a vendor).
As of yesterday I'm germinating three plates with spores taken from spore prints from my first grow and I hope that maybe I'll get a different result from those spores but I'm starting to feel like I'm losing my mind here. What is likely my problem? Shitty genetics? Shitty samples? Shitty agar? Something else? I need to figure this out quickly as petri dishes are hard for me to get and very expensive to boot!
Thanks for reading.
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SingularFusion


Registered: 10/31/18
Posts: 1,217
Last seen: 4 hours, 43 minutes
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Re: Issues with my hot pour agar technique [Re: tijuanataximan]
#27031229 - 11/10/20 08:35 AM (3 years, 3 months ago) |
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I don't know why the poor growth but feel it's gotta be recipe... if you noc them with syringes, I feel like something will grow, good or bad, unless the media is fucked to start with. Open a clean plate and let it sit open and exposed in plain air for 30 seconds, if nothing still grows, your media is junk. I reckon that's the easiest test to do, start ruling out factors one by one
the condensation: try a couple of degrees lower. The actual point at which it sets will depend on brand of agar, strength of your agar mix and what other ingredients went into it IME. Pour as close as you can get away with to the point where it sets. The last plate you pour you should basically be having agar already go slushy in the bottle
stack the plates while cooling if you aren't doing that already
don't pour agar in a cold room, condensation is all a feature of temp differential
dunno how much of that you are already doing but stick at it man you will get it nailed
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mushboy
modboy



Registered: 04/24/05
Posts: 32,389
Loc: where?
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Re: Issues with my hot pour agar technique [Re: SingularFusion]
#27031248 - 11/10/20 08:52 AM (3 years, 3 months ago) |
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Try something basic like brf agar to compare.
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tijuanataximan
Stranger

Registered: 07/09/20
Posts: 32
Loc: cananananada
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Re: Issues with my hot pour agar technique [Re: mushboy]
#27031348 - 11/10/20 10:05 AM (3 years, 3 months ago) |
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Quote:
Nef said: dunno how much of that you are already doing but stick at it man you will get it nailed
Pretty much all of it (or as close as I can get). I'm currently in the process of elim right now in regards to my media mixture. The reason I thought that the spores may be the issue is because I have no idea how old they are or how degraded they are. I'm waiting for more media to cool right now so i'll try pouring this one at an even lower temp to see if I can do it before it solidifies.
Quote:
mushboy said: Try something basic like brf agar to compare.
I will! Once I get the chance i'll run to the store to buy some BRF. I do happen to have organic rye flour handy so i'm going to whip up some media using that right now and then the BRF later.
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