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Invisiblej3rk0ff
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what is the best way to expand a isolate?
    #18867449 - 09/20/13 12:25 PM (10 years, 5 months ago)

im trying to expand a isolate and was thinking of putting the wedge into a grain jar letting it fully colonize then trying Frank's tek here to make 24 agar jars and a GLC found here for 12 master jars and lc

is the a good way of going about it?


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InvisibleFooMan
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Re: what is the best way to expand a isolate? [Re: j3rk0ff]
    #18867574 - 09/20/13 12:44 PM (10 years, 5 months ago)

agar plate=>grain jar=>agar plate=>LC=>grain jar

seems like a lot of unnecessary steps to me :shrug:


If you have an isolate on agar, there is no need to move it to grain if you plan on going back to agar. Take some small wedges from the isolate you have and transfer them to more plates. Use the wedges from those plates to inoculate your grain jars. You could also inoculate a single grain jar with an agar wedge and use that as your master grain jar which could be used to do grain-to-grain transfers to several more jars.


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Invisiblej3rk0ff
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Re: what is the best way to expand a isolate? [Re: FooMan]
    #18867673 - 09/20/13 01:04 PM (10 years, 5 months ago)

I only have 1 wedge so trying to figure out best choices from there.

if i prep my 24 agar jars and put the wedge to 1 jar then put the other 23 jars in the fridge til the one jar is done then knock up the other 23 would that work?

how long does agar hold up in a fridge?

main reason i was thinking the other way would be better is because id get 24 agar jars and 1 glc for the 12 master jars at once, but im also here to learn and gain knowledge so that is why i ask 1st.


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want to grow like a Boss
great links:
How Frank gets shit done
TL: HOW I DO EVERYTHING

Edited by j3rk0ff (09/20/13 01:30 PM)

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OfflineMush4Brains
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Re: what is the best way to expand a isolate? [Re: j3rk0ff]
    #18867825 - 09/20/13 01:39 PM (10 years, 5 months ago)

You don't knock up 23 jars with the same isolate.

Once you go from grain to agar, you transfer until you have an isolate, then you use that isolate to knock up a master jar, and then you G2G with that.

Or, since you're doing no-pour jars, assuming you have a SHIP and a GE filter, and don't want to do G2G, you can make a mycelium water syringe from your isolate plate.

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Invisiblej3rk0ff
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Re: what is the best way to expand a isolate? [Re: Mush4Brains]
    #18867887 - 09/20/13 01:58 PM (10 years, 5 months ago)

what is the reason not to have 23 isolate agar jars? degrade lost of potency from 1 isolate?

i want to do g2g i just was trying to get  24 isolate + have glc for 12 master jars to g2g then id have 24 isolates for back up to keep doing agar to g2g.

to me this makes since but do not quite understand why this would be bad.

guess i could do what your speaking of
wedge>agar>agar slurry>24 agar jars
agar>12 grain jars>g2g

this correct?

ill be using this tek just 1/2pint jars


--------------------
want to grow like a Boss
great links:
How Frank gets shit done
TL: HOW I DO EVERYTHING

Edited by j3rk0ff (09/20/13 02:09 PM)

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Invisiblebulkgrownoob
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Re: what is the best way to expand a isolate? [Re: j3rk0ff]
    #18867963 - 09/20/13 02:17 PM (10 years, 5 months ago)

If what you're looking to do is preserve your isolate to use it for a long time then you need to look into culture slants. Otherwise agar to grain for a few master jars and then g2g those

Edited by bulkgrownoob (09/20/13 02:18 PM)

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Invisiblej3rk0ff
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Re: what is the best way to expand a isolate? [Re: bulkgrownoob]
    #18868362 - 09/20/13 03:59 PM (10 years, 5 months ago)

Quote:

That's a little like asking how many miles can you drive a red car before it runs out of gas.

You'll have to quantify the question if you want more than speculation.  There's a huge difference in mycelium expansion between grain to grain transfers with 1/2 pint jars and the same thing with quart jars.  If one is using filter patch bags with 5 quarts of grains in each, the mycelium must expand much more on each transfer, thus you can't just count the number of transfers, but the actual amount of cell division.

Time also plays a big role.  Aged mycelium will not expand as far as young mycelium.

As I've posted before, I use fairly young cell lines, and expand with up to 5 grain to grain transfers with large filter patch bags before any degradation sets in, and then there's still enough energy left for that mycelium to colonize the bulk substrate and fruit.  In other words, one petri dish of mycelium can easily be expanded to enough grain spawn to fill a small warehouse if you do it fast.
RR




found some good info


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want to grow like a Boss
great links:
How Frank gets shit done
TL: HOW I DO EVERYTHING

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InvisibleSpitballJedi
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Re: what is the best way to expand a isolate? [Re: j3rk0ff]
    #18868378 - 09/20/13 04:01 PM (10 years, 5 months ago)

You don't need that many agar jars at one time.

Take a rice size sample from your agar wedge and transfer it to another agar jar or plate. Now you have a new jar and still have the master wedge.

Once colonized, use the new jar/plate to inoculate 1 or more grain jars and then G2G from there.

every time you need to create a new master grain jar, just repeat this process.

Like mentioned above, look in to slants for long term preservation of a culture.

Technically not a slant, but here's a master culture I stored in a jar. It was in my mini-fridge since February. I took it out 2-3 weeks ago and sat it on a table and forgot about it. So that's 6 months in storage and 2-3ish weeks in less than ideal conditions. I would say this is a fruiter.


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Invisiblej3rk0ff
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Re: what is the best way to expand a isolate? [Re: SpitballJedi]
    #18868944 - 09/20/13 06:16 PM (10 years, 5 months ago)

nice it even fruited. that is the type jar i got setup for. do you use wood in them? like the slants?


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want to grow like a Boss
great links:
How Frank gets shit done
TL: HOW I DO EVERYTHING

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InvisibleSpitballJedi
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Re: what is the best way to expand a isolate? [Re: j3rk0ff]
    #18868950 - 09/20/13 06:19 PM (10 years, 5 months ago)

Yes. I got some tongue depressors from the pharmacy and broke them up.


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Invisiblej3rk0ff
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Re: what is the best way to expand a isolate? [Re: SpitballJedi]
    #18868989 - 09/20/13 06:33 PM (10 years, 5 months ago)

thanks:thumbup:


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want to grow like a Boss
great links:
How Frank gets shit done
TL: HOW I DO EVERYTHING

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