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Offlinepolishfarmer
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How does this LC look ?
    #23821899 - 11/11/16 04:35 AM (7 years, 2 months ago)

How does the LC look ?

This was inoculated on 10/24. been about 2 1/2 weeks.
I don't remember being so much debris on the bottom before the inoculation. Is that the myc sitting down there? also there is a tiny piece floating at the top.

I'm 99% sure its not a contam because its not spreading and I have another jar with contams floating at the top and its spreading rapidly. It also looks like mycelium to me.

these 3 pics are of the same jar

Thanks for the input







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Offlineenlightenment
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Re: How does this LC look ? [Re: polishfarmer]
    #23821908 - 11/11/16 04:45 AM (7 years, 2 months ago)

Have you made this LC with a spore syringe?


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Offlinepolishfarmer
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Re: How does this LC look ? [Re: enlightenment]
    #23821923 - 11/11/16 04:56 AM (7 years, 2 months ago)

yes I used a spore syringe.

The LC was made with honey and I put a bit of nutritional yeast and gypsum in there aswell.


Edited by polishfarmer (11/11/16 04:58 AM)


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Offlineenlightenment
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Re: How does this LC look ? [Re: polishfarmer]
    #23821926 - 11/11/16 04:57 AM (7 years, 2 months ago)

You can never say if it is clean until you tested it.
Spores to LC is always a bad idea.

Thats how my LC looks like:



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Offlinetump
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Re: How does this LC look ? [Re: enlightenment]
    #23821928 - 11/11/16 04:58 AM (7 years, 2 months ago)

Test on a agar pasty plate. But it looks screwed to me what was the sugar content.


Edited by tump (11/11/16 05:00 AM)


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Offlinepolishfarmer
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Re: How does this LC look ? [Re: enlightenment]
    #23821930 - 11/11/16 04:59 AM (7 years, 2 months ago)

if you dont use spores how else do you start an LC ?

How did you start yours ?


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Offlinepolishfarmer
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Re: How does this LC look ? [Re: polishfarmer]
    #23821935 - 11/11/16 05:03 AM (7 years, 2 months ago)

here it is next to the one that spoiled





not much stuff in the bottom of the bad one on the left...

they all had similar amounts of debris from the beginning


Edited by polishfarmer (11/11/16 05:04 AM)


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Offlinetump
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Re: How does this LC look ? [Re: polishfarmer]
    #23821936 - 11/11/16 05:03 AM (7 years, 2 months ago)

Best way is agar wedge for clean cultures. Besides that the highest chance of success is colonized grain dumped in one like grain to lc. But thats still risky form left over endo spores on the grain.


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Offlinepolishfarmer
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Re: How does this LC look ? [Re: tump]
    #23821940 - 11/11/16 05:06 AM (7 years, 2 months ago)

for some reason all my petri dishes got contaminated so I decided to do an LC because i could use a sterile syringe... I have a flow hood and everything.. but I'm working out of my bedroom so I dunno if that is fucking my shit up..


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Offlineenlightenment
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Re: How does this LC look ? [Re: enlightenment]
    #23821943 - 11/11/16 05:06 AM (7 years, 2 months ago)

Spores to LC is crap. Doesn't matter how the spores were inoculated. Spore syringes are never 100% sterile.

tump grain to LC is bad advice...

A good LC is made from a colonized agar piece in front of a flow hood or in a SAB at least. G2G is more noob friendly than LC.
When i write noob friendly i don't mean that it is disadvantaged to LC. G2G done right is even faster than LC IME.


Edited by enlightenment (11/11/16 05:13 AM)


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Offlinetump
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Re: How does this LC look ? [Re: enlightenment]
    #23821950 - 11/11/16 05:15 AM (7 years, 2 months ago)

I said agar wedge first. If you have a flow hood what are you doing wrong. Transfer until clean on plates.


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Offlinepolishfarmer
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Re: How does this LC look ? [Re: enlightenment]
    #23821951 - 11/11/16 05:15 AM (7 years, 2 months ago)

you think its ok to run my flow hood in my bedroom. its kinda cluttered in here andI'm wondering  wtf becasue im still getting contams in my petri dishes.

I run the flow hood, use alcohol and flame sterilize and everything..

but now that i think of it i started my agar with spore syringes anyway...

how do you inoculate agar without a syringe ?


Edited by polishfarmer (11/11/16 05:15 AM)


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Offlinepolishfarmer
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Re: How does this LC look ? [Re: tump]
    #23821955 - 11/11/16 05:18 AM (7 years, 2 months ago)

is it ok to open a petri dish with contams and transfer a piece of the good myc out of there ?

i didnt want to open any of the contaminated dishes because i didnt want mold spores flying everywhere

i guess if I'm infront of the flow hood it shouldn't matter right ?


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Offlineenlightenment
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Re: How does this LC look ? [Re: polishfarmer]
    #23821958 - 11/11/16 05:19 AM (7 years, 2 months ago)

run the flow hood for an hour before you start to work.
Agar is inoculated by syringes or spore prints with a sterile cotton swab. The difference between LC and agar is that you can see contaminations much easier. You also can transfer healthy mycelium away from the contaminated parts.
If a petri dish has mold on it that already started to sporulate you should not transfer in front of your flow hood. It would spread the mold spores everywhere.


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Offlinepolishfarmer
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Re: How does this LC look ? [Re: enlightenment]
    #23821967 - 11/11/16 05:23 AM (7 years, 2 months ago)

how do you tell when it starts to sporulate ?

thanks for all the input btw.


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Offlineenlightenment
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Re: How does this LC look ? [Re: polishfarmer]
    #23821973 - 11/11/16 05:26 AM (7 years, 2 months ago)

Mold turns from white or slightly grey to green or black (the most common mold varieties).

the white part on the edge is short before sporulating:


sporulating mold:


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InvisibleMad Season
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Re: How does this LC look ? [Re: polishfarmer]
    #23822190 - 11/11/16 07:34 AM (7 years, 2 months ago)

Quote:

polishfarmer said:
for some reason all my petri dishes got contaminated so I decided to do an LC because i could use a sterile syringe... I have a flow hood and everything.. but I'm working out of my bedroom so I dunno if that is fucking my shit up..



Don't mean to be a downer but why would a different liquid media produce different results when you can't get clean on the first type of liquid media? Is this post transfer you get contaminations, or initial inoculation?


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OfflineBuster_Brown
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Re: How does this LC look ? [Re: polishfarmer]
    #23822258 - 11/11/16 08:14 AM (7 years, 2 months ago)

Quote:

polishfarmer said:


I...use alcohol...




The CDC on alcohol:

"...alcohols are rapidly bactericidal rather than bacteriostatic against vegetative forms of bacteria; they also are tuberculocidal, fungicidal, and virucidal but do not destroy bacterial spores."


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