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InvisibleKalistis
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Registered: 09/06/15
Posts: 2,265
Wedge to Oats and MS to Oats Q's
    #22398754 - 10/18/15 05:04 PM (8 years, 3 months ago)

1. I accidentally boiled my oats for 65 minutes instead of the 45 minutes I did the first time. I noticed that there are more busted grains than last time and that they appear a little dry. I still threw them into the PC for 90 minutes. Do you think this will affect colonization?

2. I did my best to tilt my quart jars while I transferred my wedges of Argentina agar to my oats 5 days ago, but it looks like all of them still ended up on top. They have grown like crazy but only on the top of my oats. Do I shake now or continue to wait?



3. All of my MS to oat jars look pretty god. The B= seems to be moving slower than my Argentina jars. One of the B+ jars looks weird. I am wondering if this is a contaminate or just a product of me over cooking my jars?

Healthy jar:



Jar in question:



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OfflinePsilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
Re: Wedge to Oats and MS to Oats Q's [Re: Kalistis]
    #22399554 - 10/18/15 07:17 PM (8 years, 3 months ago)

Quote:

Caelistis said:
1. I accidentally boiled my oats for 65 minutes instead of the 45 minutes I did the first time. I noticed that there are more busted grains than last time and that they appear a little dry. I still threw them into the PC for 90 minutes. Do you think this will affect colonization?



You'll be fine, those oats look hydrated enough. Colonization times may be slower depending on the strength of the culture you're using. Genetics is everything.


Quote:

Caelistis said:
2. I did my best to tilt my quart jars while I transferred my wedges of Argentina agar to my oats 5 days ago, but it looks like all of them still ended up on top. They have grown like crazy but only on the top of my oats. Do I shake now or continue to wait?



Why tilt the jars? Just drop the wedge in, close the lid tight, and shake it up good to re-distribute it amongst the grains.


Quote:

Caelistis said:
3. All of my MS to oat jars look pretty god. The B= seems to be moving slower than my Argentina jars. One of the B+ jars looks weird. I am wondering if this is a contaminate or just a product of me over cooking my jars?



I hate to be the bearer of bad news, but I would say that all your jars have some sort of contam. :sad:

That mycelium just looks soo light and whispy, very similar to cobweb mold.

How was your sterile procedures while using your SAB. Did you spray down the walls of the SAB and let it sit for a couple of minutes before doing your work in there? Did you remember not to let your hand hover over the exposed jar while transferring a wedge or cutting into your agar?

If you haven't already, I would take a quick look at Mad Season's sterile techniques, it helped me out a lot when I was unsure about how to go about making transfers properly inside the SAB: http://www.shroomery.org/forums/showflat.php/Number/21889950/fpart/1/vc/1


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InvisibleKalistis
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Registered: 09/06/15
Posts: 2,265
Re: Wedge to Oats and MS to Oats Q's [Re: Psilosoulful]
    #22399623 - 10/18/15 07:30 PM (8 years, 3 months ago)

I'm not sure if I was clear. I transferred 6 wedges to 6 jars of oats. I did not shake immediately after inoculation based off some advice I got saying to wait, but to tilt the jars so the wedge got buried. They did not get buried and are sitting at the top of the jar. They have since colonized the top layer and have infiltrated a little below. Should I shake them now?

The other 16 jars are all MS to oats and they really look the same as my last two batches of MS to oats. I think they are wispy because they are MS.

I used a SAB, I sprayed my SAB before working in it, I did not let my hand hover over the plates or jars, and transfer took less than 30 seconds.

I followed Mad's sterile tek the first time I worked with MS to agar. Very helpful. Very few of my plates showed contaminants.


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InvisibleLocN9ne
ɢᄋᄋd ԲᄋЯ ᄁᄋȚᅢΙᄁɢ ᄂᄋ₩ᄂΙԲᄐ
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Registered: 04/17/15
Posts: 7,076
Loc: to the brain Flag
Re: Wedge to Oats and MS to Oats Q's [Re: Kalistis]
    #22399647 - 10/18/15 07:35 PM (8 years, 3 months ago)

I don't know what that black shit is in the bottom of that jar, but they all look healthy to my besides that... if it was cobweb, which I doubt you would know by now...


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InvisibleKalistis
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Registered: 09/06/15
Posts: 2,265
Re: Wedge to Oats and MS to Oats Q's [Re: LocN9ne]
    #22399658 - 10/18/15 07:38 PM (8 years, 3 months ago)

It's only one jar that has the black shit. I am only letting it continue because I am curious as to what will come of the jar. I think it is safe to keep it with the other jars since it is sealed. I am using SFD's instead of polyfil or micropore, so I know that it is isolated.


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OfflinePsilosoulful

Registered: 09/05/14
Posts: 7,205
Last seen: 1 year, 1 month
Re: Wedge to Oats and MS to Oats Q's [Re: Kalistis]
    #22399674 - 10/18/15 07:41 PM (8 years, 3 months ago)

Quote:

Caelistis said:
I'm not sure if I was clear. I transferred 6 wedges to 6 jars of oats. I did not shake immediately after inoculation based off some advice I got saying to wait, but to tilt the jars so the wedge got buried. They did not get buried and are sitting at the top of the jar. They have since colonized the top layer and have infiltrated a little below. Should I shake them now?



That advice was wrong. When transferring agar wedges, shake immediately afterwards.

Only with MS inoculations, you shouldn't shake the jars, because you want the spores to stay close together to germinate quickly.


Quote:

Caelistis said:
The other 16 jars are all MS to oats and they really look the same as my last two batches of MS to oats. I think they are wispy because they are MS.



:justno:



Quote:

Caelistis said:
I used a SAB, I sprayed my SAB before working in it, I did not let my hand hover over the plates or jars, and transfer took less than 30 seconds.



:billymaythumbup:



Quote:

Caelistis said:
I followed Mad's sterile tek the first time I worked with MS to agar. Very helpful. Very few of my plates showed contaminants.



Awesome, it really is a great write-up! :awesomenod:


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Invisiblewowimflabbergasted
supercalifragilistic
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Registered: 07/16/12
Posts: 18,918
Re: Wedge to Oats and MS to Oats Q's [Re: Psilosoulful]
    #22399703 - 10/18/15 07:46 PM (8 years, 3 months ago)

I've had jars have that black burnt shit on the bottoms before. It doesn't come off easy but it's never been a problem I don't know if it's from the PC cycle or what. :shrug:


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