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OfflineChurchill_85
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SP to Agar (Advice Please)
    #27012763 - 10/30/20 07:33 PM (3 years, 3 months ago)

First try Spore print A+ to Agar (LMA 10g/10g/500 ml + food coloring) in a SAB on day Oct 24, did three 60 x 15 plates:







(Best pictures I could take...., the black spots are something from the inoculation loop, its made from tungsten and when its was flamed released this black substance... I should tested flaming before the procedure...)


I read that the ideal is to transfer fast right? The plate I has the most formed mycelium, is it possible to transfer already? Should I wait more?

Is the standard procedure:
- SP to Agar;
- Transfers until clean; (maybe 3 transfers?)
- Agar to 1 master grain jar;
- Master grain jar G2G for another jars;
- Spawn on minitub;
- Harvest and check for flush quality as potency etc (before eating get a tissue and clone?)
- Restart cycle


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OfflineSold Out Online
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Re: SP to Agar (Advice Please) [Re: Churchill_85]
    #27012866 - 10/30/20 08:33 PM (3 years, 3 months ago)

You’re six days ahead of me. I was thinking to go from agar to multiple (10-12) grain jars, but if one master and G2G somehow works better I’d love to know!


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OfflineChurchill_85
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Re: SP to Agar (Advice Please) [Re: Sold Out Online]
    #27012883 - 10/30/20 08:45 PM (3 years, 3 months ago)

Well I'm assuming that when you isolate you are looking for the minimal genetic variation, I know to have a monoculture takes several transfers and its not necessary but I imagine if you use several agar plates from different sources you will have also several mycelium types growing together, many less than a MSS to grain, but still more. Also not sure if just one 60 agar plate can be enough to G2G like 10 jars.

Because that i assume its better to make from 1 agar plate just 1 master jar, then reducing the genetic variation. Not sure if my logic is correct...

My first grow was from a MSS to grain and I got a very low potency flush, i'm kinda traumatized now =]. My main concern now its to find a good potency genetic and a way to keep it stored somehow.


Edited by Churchill_85 (10/30/20 08:47 PM)


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OfflineChurchill_85
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Re: SP to Agar (Advice Please) [Re: Churchill_85]
    #27028761 - 11/08/20 02:49 PM (3 years, 3 months ago)

Here is an update of T1 done on Nov01:







T1A III seems to have a bacterial white circle.

- Is it possible to transfer already or need to wait as the mycelium strengthen?

Still trying to figure out some points as:

- What is that transparent mycelium, any points why that happens?

- I buy 10 dishes sleeves and when its opened i think its a good idea to pour all of them. How to proper store unused (non inoculated) dishes? Put it again on the sleeve close with plastic tape but wrapped or unwrapped? What temperature is the best?

- To pour 10 plates a small amount of agar is necessary, but i believe its best to prepare like 400 ml agar (controlling 47°C on 400 ml seems easier than on 100 ml to proper pour), pour the 10 plates and store the leftover bottle on the fridge then microwave then PC it again for the next pour. Is that ok or the agar will deteriorate?

- At some point I will have to decide which one of the plates will go to grain, is it a good idea to transfer 1 more time for safety if the master jar contam? What do you guys do with the dishes that didnt go for grain, trow away or keep it?

- Is it possible to prepare 11 jars, use 1 for the master and keep the other 10 for G2G? Or its better to prepare just 1 jar and when its ready prepare the other 10?

- For safety is better to have 2 master jar from 1 dish? I dont think is recommended mixing different dishes to make a master right?


Edited by Churchill_85 (11/08/20 02:55 PM)


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Offlineverum subsequentis
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Re: SP to Agar (Advice Please) [Re: Churchill_85] * 2
    #27028807 - 11/08/20 03:19 PM (3 years, 3 months ago)

- What is that transparent mycelium, any points why that happens?

I believe it can happen for many different reasons. In your particular case I'd suggest just making another transfer and seeing how she turns out. Germination plates and the next plate or two can be a little strange.

- I buy 10 dishes sleeves and when its opened i think its a good idea to pour all of them. How to proper store unused (non inoculated) dishes? Put it again on the sleeve close with plastic tape but wrapped or unwrapped? What temperature is the best?

You can store many ways. All that matters is that they are sealed as to not contaminate. I prefer wrapping them with glad cling wrap. I cut a piece off the roll that is just a little wider than the edge of the dish and double wrap the dishes. Storing at room temp is fine. What you want avoid are dramatic temperature fluctuations. Temp flux draw moisture out of the agar and condensate your plates. This is bad for many reasons.


- To pour 10 plates a small amount of agar is necessary, but i believe its best to prepare like 400 ml agar (controlling 47°C on 400 ml seems easier than on 100 ml to proper pour), pour the 10 plates and store the leftover bottle on the fridge then microwave then PC it again for the next pour. Is that ok or the agar will deteriorate?

I prefer to make a bunch and pour it all in one go. I do one liter to 50-60 plates depending on how thick i pour. You can do a little and then reheat the agar if you prefer. I find it to be less work to just use what is made though. I'd skip the microwave and just PC if i were to do it your way though. Why microwave? The PC will take care of that.

Also, you don't have to PC again if you haven't contaminated your agar. Done correctly you could just microwave it and then pour again. Careful or you'll end up with a ginormous mess and some possible burns. That shit can get crazy fast in a microwave.

I'd strongly suggest just pouring it all in one session though. I've done it enough to not really run into the problem anymore but you can just eyeball the left over agar and decide whether or not to start more dishes. pouring the last few a little thinner or a little thicker you can always end up using all the plates you've opened. If you end up with a few plates opened but not poured, just wrap them like the rest and use in your next session.

- At some point I will have to decide which one of the plates will go to grain, is it a good idea to transfer 1 more time for safety if the master jar contam? What do you guys do with the dishes that didnt go for grain, trow away or keep it?

Depends on how many dishes i have of a particular culture laying around. In your case I'd keep it in case you need it or in case it pins later and you want a quick clone.

Always transfer a small piece to another plate before committing the plate to a jar as to be sure you don't lose the culture.

- Is it possible to prepare 11 jars, use 1 for the master and keep the other 10 for G2G? Or its better to prepare just 1 jar and when its ready prepare the other 10?

Meaning, keep the others uninocculated and save for later use? You can but it is not recommended. Better to use them as soon ass possible and make more when you need them.

- For safety is better to have 2 master jar from 1 dish? I dont think is recommended mixing different dishes to make a master right?

It is not a good idea to use two plates for one jar. Specially for a noob. It's hard sometimes for even experienced cultivators to spot sneaky contams on plates.

As a noob it is always safer to make a few more of something than you need. Two masters is safer than one, for sure.


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OfflineChurchill_85
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Re: SP to Agar (Advice Please) [Re: verum subsequentis]
    #27028823 - 11/08/20 03:29 PM (3 years, 3 months ago)

Thank you for the repply!


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Offlineverum subsequentis
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Re: SP to Agar (Advice Please) [Re: Churchill_85]
    #27028960 - 11/08/20 05:13 PM (3 years, 3 months ago)

My pleasure


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