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OfflineGermanShaman
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Is this contam in my LC?
    #27107342 - 12/25/20 12:08 PM (3 years, 1 month ago)

Apologies if this seems a little obvious, I just want to make sure that I dont throw out good LC.

Take a look at these pictures and let me know if this is all contam please. They were all inoculated from the same agar dish.

LC1a-c  is 4% Honey solution

LC2a-c is 4% Honey Solution

LC3a-c is 4% LME solution.

They were all PC sterilised prior to inoculation and have been kept at around 22-24 Dec C.

On LC 1 & 2 there is clearly blue mould on the surface.

LC3 seems a bit frothy but no signs of mould.


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OfflineGermanShaman
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Re: Is this contam in my LC? [Re: GermanShaman]
    #27107346 - 12/25/20 12:09 PM (3 years, 1 month ago)

Here are the pics for LC 1a - c



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OfflineGermanShaman
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Re: Is this contam in my LC? [Re: GermanShaman]
    #27107347 - 12/25/20 12:10 PM (3 years, 1 month ago)

Here are the pics for LC 2a - b



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OfflineGermanShaman
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Re: Is this contam in my LC? [Re: GermanShaman]
    #27107351 - 12/25/20 12:11 PM (3 years, 1 month ago)

Here are the pics for LC 3a - b



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OfflineHarry Manbach
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Re: Is this contam in my LC? [Re: GermanShaman]
    #27107355 - 12/25/20 12:13 PM (3 years, 1 month ago)

Toss it!

I'd definitely suggest investing in self healing injection ports ..as well as hydrophobic syringe filters,especially for lc lids! It'd help mitigate contamination MUCH more than micropore!


Edited by Harry Manbach (12/25/20 12:14 PM)


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InvisibleLemgrub
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Re: Is this contam in my LC? [Re: Harry Manbach]
    #27107363 - 12/25/20 12:16 PM (3 years, 1 month ago)

All bad:thumbdown:


--------------------
We getting that pirate bay, alien shishkebab cordycep money


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OfflineGermanShaman
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Re: Is this contam in my LC? [Re: Lemgrub]
    #27107581 - 12/25/20 03:49 PM (3 years, 1 month ago)

OK. I will toss these.

I do have some lids with self healing injection ports that I made with high temp silicone sealant.  The thing is, how do I inoculate those using culture from Agar?

Or do I use the agar to test the syringe and then use the syringe to inoculate the culture?


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Invisiblemicelio
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Re: Is this contam in my LC? [Re: GermanShaman]
    #27107673 - 12/25/20 05:14 PM (3 years, 1 month ago)



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OfflineGermanShaman
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Re: Is this contam in my LC? [Re: micelio]
    #27108213 - 12/26/20 03:24 AM (3 years, 1 month ago)

thank you for those links micelio.

very interesting reading.

I have been using 4% solutions but these guys are using far more dilute solutions, around 0.2%.  Am I going over board?  Maybe I should try a lower concentration of LME or Honey.


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Invisiblemicelio
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Re: Is this contam in my LC? [Re: GermanShaman]
    #27108900 - 12/26/20 12:42 PM (3 years, 1 month ago)

Rule one: Is the KISS Rule. Keep It simple stupid.


This Is the style jar I been using.

The jar is 6" tall (the length of a dollar bill)

The jar has a 2" dia. opening.

The jars had salsa in them, I like salsa with my corn chips so I'm getting a deal.. Ha



I toss out all my filters and injection ports, too much clutter.

Just one 6/64" drill hole so my 16 gauge needle just fits snug.

I cut a piece of masking tape and cover the hole.

Before I pc the jars I loosen all the jar lids a little.

After pc cycle I tighten the lids while jars are hot. Then wait for the security button to click on the jar lids as they cool down and pull a vacuum.

The jars are now under a vacuum waiting to be inoculated. After the jars are noc. up I cut a piece of masking tape and cover the hole.

One important note: If you jab the hole with the needle while jar Is under a vacuum You'll find yourself with an empty syringe real quick.
Poke a hole in the tape over the hole to let the vacuum out of the jar first.


To aspirate the lc is a simple process of placing the jar in a horizontal position keeping the hole just above the fluid level.

There should be enough air seep in through the same hole to not create a vacuum inside the jar that the needle is in. Also this should be done in a sab or in the flow. (LFH)



Edited by micelio (12/26/20 03:11 PM)


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