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eatyualive
Eat's You Alive :)



Registered: 08/17/01
Posts: 19,026
Loc: In Your Head
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Re: Constant Agar contamination? So Frustrating. [Re: gizmo1]
#26474684 - 02/07/20 08:51 PM (3 years, 11 months ago) |
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You could try micropore tape over the holes instead of poly. But I don’t think that is the issue.
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PiptheGreAtest
Aspiring Heavyweight


Registered: 07/18/16
Posts: 464
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Re: Constant Agar contamination? So Frustrating. [Re: eatyualive]
#26479845 - 02/11/20 09:41 AM (3 years, 11 months ago) |
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I appreciate all the advice, I’m going to get a cookie sheet to raise my work space. I bought some pre sterilized plates just to see if it was my technique and work space or the peeping of my agar. The bathroom is never used so area shouldn’t be a problem. The poly fill in in there really tight and I guess that would be my GE for the plates.
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Jarhead3521
Grindin’



Registered: 09/26/19
Posts: 514
Loc: Amongst the pines
Last seen: 5 months, 21 days
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Re: Constant Agar contamination? So Frustrating. [Re: PiptheGreAtest]
#26479865 - 02/11/20 09:52 AM (3 years, 11 months ago) |
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I’m going to x2 on the smooth movement in the sab, and letting things settle. Last round of petris I poured, was not smooth. Used 10 out of the 20 the following day and left the unused stack in the box....been there for a couple weeks, started to do some work last night and noticed 2 of 10 had shit growing in them, and it most definitely came from my technique that night.
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PiptheGreAtest
Aspiring Heavyweight


Registered: 07/18/16
Posts: 464
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Re: Constant Agar contamination? So Frustrating. [Re: Jarhead3521]
#26479909 - 02/11/20 10:20 AM (3 years, 11 months ago) |
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Smooth movements and fix holes got it.
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Sockadin



Registered: 01/03/10
Posts: 7,244
Last seen: 2 months, 20 days
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Re: Constant Agar contamination? So Frustrating. [Re: PiptheGreAtest]
#26479933 - 02/11/20 10:37 AM (3 years, 11 months ago) |
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Quote:
PiptheGreAtest said: I have done multiple runs of agar in patsywhtyes easy agar dishes, every time they have contaminated and failed. I’m inoculating with MS syringe in a SAB, I flame the needle with alcohol lamp. SAB is sprayed down with bleach cleaner and air sanitizer along with the room in a similar fashion. I choose to wear tyvek suit with mask and gloves all which get sprayed with disinfectant. The plates are prepared how pastsy describes and PC’d for 45 minutes. Why are these plates failing? Any advance cultivator if I could get tips on how to prevent this that would be great.

Haven't we decided that the lids can be unmodified now days. Is the poly even needed?
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PiptheGreAtest
Aspiring Heavyweight


Registered: 07/18/16
Posts: 464
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Re: Constant Agar contamination? So Frustrating. [Re: Sockadin]
#26479945 - 02/11/20 10:45 AM (3 years, 11 months ago) |
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I’m stepping away from patsy plates. I need better visibility so I got some pre sanitize and patsy plates dishes. I’m trying to isolate my problem but in further will either get glass Petrie dishes.
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InfiniteDreams


Registered: 10/25/19
Posts: 1,224
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Re: Constant Agar contamination? So Frustrating. [Re: PiptheGreAtest]
#26479966 - 02/11/20 10:59 AM (3 years, 11 months ago) |
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When you you inoculate your plates or whatever you are using, leave 1 or 2 with agar but don't inoculate them. These can be your controls. If they contaminate then you know you have an issue in your prep phase before you even get to inoculation.
If they don't contaminate but your inoculated plates do, then you know you either have issue with how you inoculate or it is just coming from the source.
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Backbone
Stranger



Registered: 01/18/20
Posts: 339
Last seen: 3 years, 8 months
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Re: Constant Agar contamination? So Frustrating. [Re: InfiniteDreams]
#26480111 - 02/11/20 12:41 PM (3 years, 11 months ago) |
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i inoculated three plates in a sab pcd my agar beforehand, sanitized before and havent got any contams in 3 days, its probably his syringe, my methods werent perfect i touched a dish and had condensation in each plate...I got the idea to wait for the air to still in my sab, but i didnt use anything to elevate my plates i just made sure i wiped the entire area of the sab, this of course still pending. I heard only a couple days are needed before signs of contam show, its been three, ill keep watching.
Edited by Backbone (02/11/20 12:49 PM)
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A.k.a
Stranger



Registered: 10/27/19
Posts: 16,782
Loc: Gaming the system
Last seen: 15 minutes, 4 seconds
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Re: Constant Agar contamination? So Frustrating. [Re: Backbone]
#26480149 - 02/11/20 01:11 PM (3 years, 11 months ago) |
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Well you’re supposed to wait for the air to settle before using it so if you haven’t been that could be a big reason.
I see some people work super high up in the box, my metal grate thing only keeps things maybe half an inch off the bottom but it’s been fine.
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LAGM2020     
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