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OfflineDeadPhan
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Eager to get started...is it ok if I transfer once from agar and move to grain?
    #18831965 - 09/12/13 10:44 AM (10 years, 4 months ago)

I kinda just want some masters ASAP. I have only made one transfer, however, to like 10 plates. If I was to make a master grain from the nicest one as is, and then, make further isolates from the others, that should work fine, no?

I mean considering they are from ms and I don't know how any if it performs anyway wouldn't it be ideal to see how they fruit and clone from there?  I'm still trying to figure out agar

From what I've heard, even seeing good sectors and making isolates from MS innocs won't tell you how they perform in fruiting.  Correct?


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Big Gulps!  Alright!  Well,  See ya later!
And if i claim to be a wise man, well, it surely means that i dont know!


Edited by DeadPhan (09/12/13 10:45 AM)


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Offlinekinkaku
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: DeadPhan]
    #18832198 - 09/12/13 12:02 PM (10 years, 4 months ago)

correct you can have an isolate that doesnt fruit although pretty rare. your idea is a pretty good one:thumbup:


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OfflineAmanita virosa
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: kinkaku]
    #18832369 - 09/12/13 12:59 PM (10 years, 4 months ago)

yes, you can.  But if you are Not going to use a single strain isolate, then you might as well use a bunch of strains and let them fight it out, since you will have no way of knowing which of the two or three strains you may have transferred onto the plate did the best at fruiting.  That said, it is pretty rare for a vigorously growing isolate to be completely sterile in my experience assumeing you are talking about cubes.  Outside the cube world, there are many extremely vigorous anamorphic strains of certain species.


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OfflineDeadPhan
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: Amanita virosa]
    #18832771 - 09/12/13 02:44 PM (10 years, 4 months ago)

Ok....on that note, I should wait for the plate to fully colonize before transferring to grain yes?  I gather I can make 4 master jars easy from one half pint agar jar?  Or can I just make one also that would colonize a lot quicker?

I have so many jars that it wouldn't be such a big deal to use whole jars worth on a grain master.  Considering the circumference of a half pint mason isn't so big anyway. What say you?


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Big Gulps!  Alright!  Well,  See ya later!
And if i claim to be a wise man, well, it surely means that i dont know!


Edited by DeadPhan (09/12/13 02:46 PM)


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InvisibleFooMan
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: DeadPhan]
    #18832820 - 09/12/13 02:59 PM (10 years, 4 months ago)

Quote:

DeadPhan said:
Ok....on that note, I should wait for the plate to fully colonize before transferring to grain yes?




Preferably you want to cut the newest mycelium from the outer edge when it starts growing. 

Quote:

I gather I can make 4 master jars easy from one half pint agar jar?




Yes. You only need a small wedge to get things going.

 
Quote:

Or can I just make one also that would colonize a lot quicker?




Sure, more wedges will give you more inoculation points. You could also drop a wedge in an LC and really multiply the myc.


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OfflineAmanita virosa
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: DeadPhan]
    #18832898 - 09/12/13 03:22 PM (10 years, 4 months ago)

Quote:

DeadPhan said:
Ok....on that note, I should wait for the plate to fully colonize before transferring to grain yes?  I gather I can make 4 master jars easy from one half pint agar jar?  Or can I just make one also that would colonize a lot quicker?

I have so many jars that it wouldn't be such a big deal to use whole jars worth on a grain master.  Considering the circumference of a half pint mason isn't so big anyway. What say you?




I like to cut the wedges very small and scoop them up en mass on the scalpel. I shoot for five or six wedges per quart jar. As foo said the more u use the faster colonization will be. One trick is carefully coaxing the wedges deep down into the grain by shaking  there is an art to it as they like to stick onto the glass  Also ideally you use a plate when it come to about one cm of the edge but before it reaches the edge  If the plate is fully ciolonized, leave the last cm edge behind. This is the most likely part to show contam.


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InvisibleHypnotoadCroaked
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: FooMan]
    #18832926 - 09/12/13 03:29 PM (10 years, 4 months ago)

The best example of how isolates act was provided by Notahacker420:

http://www.shroomery.org/forums/showflat.php/Number/18219379#18219379


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OfflineDeadPhan
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: Amanita virosa]
    #18833587 - 09/12/13 06:10 PM (10 years, 4 months ago)

Quote:

Amanita virosa said:
Quote:

DeadPhan said:
Ok....on that note, I should wait for the plate to fully colonize before transferring to grain yes?  I gather I can make 4 master jars easy from one half pint agar jar?  Or can I just make one also that would colonize a lot quicker?

I have so many jars that it wouldn't be such a big deal to use whole jars worth on a grain master.  Considering the circumference of a half pint mason isn't so big anyway. What say you?




I like to cut the wedges very small and scoop them up en mass on the scalpel. I shoot for five or six wedges per quart jar. As foo said the more u use the faster colonization will be. One trick is carefully coaxing the wedges deep down into the grain by shaking  there is an art to it as they like to stick onto the glass  Also ideally you use a plate when it come to about one cm of the edge but before it reaches the edge  If the plate is fully ciolonized, leave the last cm edge behind. This is the most likely part to show contam.



Do you have to flame scalpel between cutting on the same plate?    How many quart jars do you tend to make from one plate?  What kind of plates do you work with?  I'm working with half pint mason jars.


--------------------
 


Big Gulps!  Alright!  Well,  See ya later!
And if i claim to be a wise man, well, it surely means that i dont know!


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InvisibleSpitballJedi
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: DeadPhan]
    #18833655 - 09/12/13 06:23 PM (10 years, 4 months ago)

Since my PC happens to hold 7 jars, I use 1 plate for 7 jars.

...and petri dishes are way easier to use, in my opinion.


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OfflineDeadPhan
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: SpitballJedi]
    #18837382 - 09/13/13 03:22 PM (10 years, 4 months ago)

So basically, I've made a bunch of agar jars from transfers that were made from MS syringes.

Is there really any point to make transfers from these jars considering, isolating any further could be in vain?  In that, y not just make masters from these and then see what performs the best and clone from there.

I don't see the point in wasting propane or agar for that matter just to further isolate from multispore without even knowing how they will perform.

A I correct in this assumption?

Most of these new dishes that I transfers from original MS inoculation are looking really good.  I figure I just get some grains ready to nocc up within the next couple days and go from there.

What say you?


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Big Gulps!  Alright!  Well,  See ya later!
And if i claim to be a wise man, well, it surely means that i dont know!


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OfflineRogerRabbitM
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: DeadPhan]
    #18837410 - 09/13/13 03:30 PM (10 years, 4 months ago)

Quote:

I don't see the point in wasting propane or agar for that matter just to further isolate from multispore without even knowing how they will perform.

A I correct in this assumption?




No.

The time spent isolating strains is worth it because you can then grow any awesome strain you create for life by storing on master slants.

What you'd be doing now is simply multispore inoculation with an added vector of contamination that wouldn't have been there if you'd simply went from spores to grain.  Spend two to three weeks now to isolate the ten or so best looking sectors and then test fruit each one.  Look for the best performer(s) and then go back to those plates to transfer to future grain masters.

If you simply clone a fruit from multispore inoculation, it's likely to be made up of several strains, so you're still back to nearly square 1.
RR


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InvisibleFrankHorrigan
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: RogerRabbit]
    #18837422 - 09/13/13 03:33 PM (10 years, 4 months ago)

Quote:

DeadPhan said:
I mean considering they are from ms and I don't know how any if it performs anyway wouldn't it be ideal to see how they fruit and clone from there?  I'm still trying to figure out agar

From what I've heard, even seeing good sectors and making isolates from MS innocs won't tell you how they perform in fruiting.  Correct?




Yes and yes.

You're on the right track.

All of my finest fruiting isolates came from what you described:

spores to agar, a transfer or two, and then inoc with the resulting plates. Clone the biggest clusters and fastest growing fruits.

I call these "limited" MS grows, they are fantastic for selecting fruits to clone.

Quote:

RogerRabbit said:
If you simply clone a fruit from multispore inoculation, it's likely to be made up of several strains, so you're still back to nearly square 1.
RR




But these can be isolated further and then tested again :shrug:

I don't know about DP but I don't have the time or space or capacity to isolate all the good looking strains on an MS plate. That is a whole lot of transfers (and plates). I'd rather let the mycelium show me the best fruiters. Just my opinion I guess.


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Here is how I get things done.
You should take a look. :hehehe:


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Edited by FrankHorrigan (09/13/13 03:38 PM)


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InvisibleSpitballJedi
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: FrankHorrigan]
    #18837439 - 09/13/13 03:37 PM (10 years, 4 months ago)

That's my opinion as well


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OfflineAmanita virosa
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: DeadPhan]
    #18838316 - 09/13/13 07:20 PM (10 years, 4 months ago)

Quote:

DeadPhan said:
Quote:

Amanita virosa said:
Quote:

DeadPhan said:
Ok....on that note, I should wait for the plate to fully colonize before transferring to grain yes?  I gather I can make 4 master jars easy from one half pint agar jar?  Or can I just make one also that would colonize a lot quicker?

I have so many jars that it wouldn't be such a big deal to use whole jars worth on a grain master.  Considering the circumference of a half pint mason isn't so big anyway. What say you?




I like to cut the wedges very small and scoop them up en mass on the scalpel. I shoot for five or six wedges per quart jar. As foo said the more u use the faster colonization will be. One trick is carefully coaxing the wedges deep down into the grain by shaking  there is an art to it as they like to stick onto the glass  Also ideally you use a plate when it come to about one cm of the edge but before it reaches the edBge  If the plate is fully ciolonized, leave the last cm edge behind. This is the most likely part to show contam.



Do you have to flame scalpel between cutting on the same plate?    How many quart jars do you tend to make from one plate?  What kind of plates do you work with?  I'm working with half pint mason jars.



I make five quarts per plate. If you cut the wedges small you could do more. I use a bacinerator and I flame between jars. Only takes a sec. I use vented 90 mm plates. Make more sence than mason jars  much easier to work  with  and have a larger diameter means more wedges.  They come in sleeves of 20 presterilized and are relatively cheap. We buy them by the case of 25 though. We also sell them.  Agar is very easy once u get the hang if it. I never fuck with LC anymore. Agar is much more reliable IMO


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InvisibleVirginiaGentleman
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Re: Eager to get started...is it ok if I transfer once from agar and move to grain? [Re: Amanita virosa]
    #18838421 - 09/13/13 07:45 PM (10 years, 4 months ago)

Quote:

Amanita virosa said:
Agar is very easy once u get the hang if it. I never fuck with LC anymore. Agar is much more reliable IMO





I never quite got the hang of LCcc's:2G, wedging is hard to mess up unless your grain prep & sterile Tek sucks


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