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Offlinefractaldill
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Inoculating Grain with an Agar Wedge
    #19587433 - 02/18/14 09:32 PM (10 years, 4 months ago)

Pretty simple question. How large of an agar wedge do you cut out to inoculate a grain jar?

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OfflineStromriderM
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Re: Inoculating Grain with an Agar Wedge [Re: fractaldill]
    #19587447 - 02/18/14 09:36 PM (10 years, 4 months ago)

Instead of one big one cut up a bunch of small ones and put in there and shake them around to dispurse. That way it will colonize much faster

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Offlinefractaldill
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Re: Inoculating Grain with an Agar Wedge [Re: Stromrider]
    #19587458 - 02/18/14 09:39 PM (10 years, 4 months ago)

but how large?

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OfflineDannyDGAF
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Re: Inoculating Grain with an Agar Wedge [Re: fractaldill]
    #19587466 - 02/18/14 09:40 PM (10 years, 4 months ago)

Size of rice is great.  If it's an isolate your using, use multiple sized rice size wedges, if not an isolate, try to stick to one unless you don't care what kind of genetics you get (such as with a MS syringe)


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OfflineStromriderM
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Re: Inoculating Grain with an Agar Wedge [Re: DannyDGAF]
    #19587549 - 02/18/14 09:58 PM (10 years, 4 months ago)

I usually do 4 dime size pieces

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Offlinefractaldill
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Re: Inoculating Grain with an Agar Wedge [Re: Stromrider]
    #19587563 - 02/18/14 10:01 PM (10 years, 4 months ago)

Do you guys let it colonize the entire plate? I've been letting it colonized the entire plate and cutting that into 4 pieces. From there i inoculate 4 grain masters. should i make the pieces smaller?

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OfflineStromriderM
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Re: Inoculating Grain with an Agar Wedge [Re: fractaldill]
    #19587579 - 02/18/14 10:03 PM (10 years, 4 months ago)

Man just do whatever works for you. Their ain't no rules baby. Experiment a little and see what works best for you. That's part of the fun

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Offlinefractaldill
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Re: Inoculating Grain with an Agar Wedge [Re: Stromrider]
    #19587596 - 02/18/14 10:08 PM (10 years, 4 months ago)

Trying to be as efficient as possible :p

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Offlinesytar
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Re: Inoculating Grain with an Agar Wedge [Re: Stromrider]
    #19587609 - 02/18/14 10:12 PM (10 years, 4 months ago)

Why does RR insist on one wedge face up? I've only done a few experiments but cutting up a ton of small pieces then throwing in a decent sized chunk and shaking seems to work dramatically faster.


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OfflineStromriderM
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Re: Inoculating Grain with an Agar Wedge [Re: sytar]
    #19587616 - 02/18/14 10:14 PM (10 years, 4 months ago)

Yeah I get much faster colonization by putting a few small pieces in there instead of one big piece

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InvisibleFrankHorrigan
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Re: Inoculating Grain with an Agar Wedge [Re: Stromrider]
    #19587894 - 02/18/14 11:17 PM (10 years, 4 months ago)

I personally hate cutting multiple wedges for one jar. Each wedge is another chance for a contam to slip in.

I inoculate my master jars with just a single wedge, the size of a quarter to a half dollar. I keep them at around 78F and I shake once after 6-8 days. They never take longer than a couple weeks :shrug:


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Offlinefractaldill
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Re: Inoculating Grain with an Agar Wedge [Re: FrankHorrigan]
    #19587898 - 02/18/14 11:18 PM (10 years, 4 months ago)

And throw the rest of the plate away?

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InvisibleFrankHorrigan
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Re: Inoculating Grain with an Agar Wedge [Re: fractaldill]
    #19587918 - 02/18/14 11:23 PM (10 years, 4 months ago)

No, that is usually the size of my culture when I inoculate my jars.

If it is bigger, I will cut it in half but I will still only ever use one wedge per jar :thumbup:

And sometimes some of the mycelium does get tossed. I never go out of my way to try to "save" or use all the mycelium on every dish.

For example, here is a dish with a section I used.  The culture itself was slightly larger than a half-dollar. Half of the culture was used to inoculate one jar and then transfers were taken from the other side for further expansion. The remaining was thrown out after being photographed.



Quote:

sytar said:
Why does RR insist on one wedge face up?




I doubt he does anymore, or at least not for cubensis.


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Offlinesytar
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Re: Inoculating Grain with an Agar Wedge [Re: FrankHorrigan]
    #19588659 - 02/19/14 05:52 AM (10 years, 4 months ago)

Dunno about that. I made a thread about it in the last week and that's what he told me to do.


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OfflinePussyFart
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Re: Inoculating Grain with an Agar Wedge [Re: sytar]
    #19588661 - 02/19/14 05:56 AM (10 years, 4 months ago)

I have done tests placing the mycellium side up, down, and on it's side.

I saw no real difference in speed of growth.

I always place them face uu tho......it's more of a personal preference than anything.

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OfflineRogerRabbitM
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Re: Inoculating Grain with an Agar Wedge [Re: PussyFart]
    #19588691 - 02/19/14 06:18 AM (10 years, 4 months ago)

A wedge the size of a dime will colonize a quart jar in a week.

Never allow a petri dish to fully colonize.

Quote:

Why does RR insist on one wedge face up?




I don't insist.  I've said the mycelium jumps off slightly faster if right-side-up.  Usually, they flip as they drop so you really don't know how they'll land.  If the agar lands mycelium down, leave it.
RR


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OfflineGeneric
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Re: Inoculating Grain with an Agar Wedge [Re: RogerRabbit]
    #19589454 - 02/19/14 11:06 AM (10 years, 4 months ago)

I was wondering about this agar wedge of a clone that inoculated this rye jar.


I was using no-pour agar in a half-pint mason jar, and think I poured the agar too deep.  It's like 3/4 of an inch deep.

Mycellium only grows on the surface of agar right?  would having this much excess agar be a problem?

What is this talk about rice-grain-sized pieces being ideal for agar-to-grain transfers?  I thought more would be better...

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InvisibleFrankHorrigan
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Re: Inoculating Grain with an Agar Wedge [Re: FrankHorrigan]
    #19589478 - 02/19/14 11:17 AM (10 years, 4 months ago)

Quote:

FrankHorrigan said:
Each wedge is another chance for a contam to slip in.




As long as no contams stuck to your agar during inoculation, it will be fine.


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You should take a look. :hehehe:


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Invisible36fuckin5
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Re: Inoculating Grain with an Agar Wedge [Re: FrankHorrigan]
    #19589492 - 02/19/14 11:20 AM (10 years, 4 months ago)

I use 60mm plates, and I generally use a whole plate for a master jar. As with Frank, though, I never let my cultures touch the edge, and if they happen to hit it, I do another transfer on agar first.

A wedge the size of a grain of rice is too small to start a quart jar. It works, but it's slow as hell. Cutting multiple  wedges means your scalpel has to touch your media more times, leaving more chances for contamination to hit.


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InvisibleFrankHorrigan
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Re: Inoculating Grain with an Agar Wedge [Re: 36fuckin5]
    #19589507 - 02/19/14 11:26 AM (10 years, 4 months ago)

Quote:

36fuckin5 said:
I use 60mm plates, and I generally use a whole plate for a master jar. As with Frank, though, I never let my cultures touch the edge, and if they happen to hit it, I do another transfer on agar first.




Have you ever had an issue with the cultures hitting the edge? This is one of those things I've read a lot about on paper but never seen in practice.

I always try to use the plates before the cultures hit the edge, but when they do I just cut a bit inside the edge, leaving the outer bit of myc behind on that side.


--------------------

Yes, you can bump my old threads with a question.
Here is how I get things done.
You should take a look. :hehehe:


Frank's tips and tricks. Updated on 3/21/14
AMU- Get an answer here -AMU

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