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Agar, strain isolation and grow thread


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#1 invisibilitysyndrome

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Posted 04 February 2016 - 02:55 PM

20160204_124325.jpg 20160204_124327.jpg 20160204_124316.jpg 20160204_124301.jpg 20160204_124251.jpg 20160204_124242.jpg 20160204_124230.jpg 20160204_124212.jpg 20160204_124152.jpg 20160204_124139.jpg 20160204_123951.jpg

So here are
TLY done from MS solution first 2 plates are the original ms be for the first transfer..
AA+ from Ms solution transfered twice sofar
Last pic is golden teacher there are a few more gt but they all look relatively the same. They are on third transfer and they were done from a tissue clone.
I'm looking for any insight, advice and conversation on using agar.
This is my first successfull agar run with multiple transfers.
Mind you I've not grown any of these out so I don't know how they will produce. I guess I need some insight on how to continue and if we can tell witch ones are ready to be put to grain.
Thnx all. Cheers


Edited by Myc, 29 February 2016 - 10:25 AM.

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#2 peacefrog

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Posted 04 February 2016 - 06:50 PM

Nice agar plates and pics. All plates look like they have sectoring still going on, but the last one looks real good. I would do several transfers from that plate for sure. You may need a few more after that or you may have yourself a few pure cultures after you transfer.

If you are looking to get several pure cultures to test, you are defintely getting close. Just continue to transfer until you have complete uniform growth, then you can start testing those bad boys. That's my favorite part.

Nicely done!
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#3 Needles

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Posted 04 February 2016 - 11:33 PM

Your plates look very good for your first go at it. Even though your last plate has the best rizmorphic growth, it looks like a satellite colonie of something other than what you want. maybe grab from the leading edge of the plate second to last. It seems to have some very healthy mycelium going on. Nevertheless, you have some nice looking plates. You may also want to cool your agar without stirring or mixing once sterilized. That will help keep the agar from forming bubbles in your dishes. Nice work waiting to see more.....
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#4 invisibilitysyndrome

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Posted 05 February 2016 - 04:34 AM

Right on yall
@Peacefrog @needles.
Preciate the insight. What do you mean needles by a satellite colony other than what i want?

That last plate is one of 7 of GT that is firther along than all the others not shown its a 3rd transfer from a tissue clone. So basicly a tissue sample grown on agar and then sectored and transfered 3 more times.
The other plates are 2nd transfers from multi spore solution . #3 4 5 6 are tly . Next 4 are aa+

So yall feel like that last one is almost there?
Far as the bubbles go ya i shook the agar up cause it was cool and hard so it was reheated on the stove and just befor i went to pour it i seen some chunks so i shook it and promptly realized what i did .

I have a bunch more plates theses were best most recent.
Soon enough ill go to master slants after they are grown out of course..
Thnx for the insight...
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#5 peacefrog

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Posted 05 February 2016 - 06:37 AM

Sorry, Needles, I'm not sure what you are saying either.

That plate is very close to uniform rhizomorphic growth, and has the least amount of sectoring compared to the other transferred plates.

Invisibility, I am pulling up a chair for this one and look forward to your results. After you get all isolated are you going to test each in larger grows? Or are you thinking of doing a bunch in mini grows? I personally like the mini option on cased grain for cubes, as it's much faster and you can test several side by side at the same time with little space.

#6 invisibilitysyndrome

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Posted 05 February 2016 - 07:14 AM

Sorry, Needles, I'm not sure what you are saying either.
That plate is very close to uniform rhizomorphic growth, and has the least amount of sectoring compared to the other transferred plates.
Invisibility, I am pulling up a chair for this one and look forward to your results. After you get all isolated are you going to test each in larger grows? Or are you thinking of doing a bunch in mini grows? I personally like the mini option on cased grain for cubes, as it's much faster and you can test several side by side at the same time with little space.

Ya im thinking smaller test runs till things are going smooth. Ive been working these plates fer a while that ive not really planed the next phase yet but i like the small shoeboxes cause i have those already but may go smaller like little tupperware/glad things.
Im lookin forward to seeing the diff in the GT compared to MS g2g
This is the first ive grown other than GT got the aa tly prints and been waiting to get em on agar.
It was deff time cause they were getting weak.

#7 Needles

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Posted 05 February 2016 - 11:31 AM

I was looking at the growth in the one o'clock position on the edge of the last plate. If it was a contaminant of some sort it may compromise future transfers.
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#8 invisibilitysyndrome

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Posted 05 February 2016 - 12:31 PM

I was looking at the growth in the one o'clock position on the edge of the last plate. If it was a contaminant of some sort it may compromise future transfers.


Ill have to go have a look see. I have several of these GT plates that all look basicly the same. One way or another ill get a pure culture.
Thnx fer all the insight yall. Ill update as it progresses.
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#9 peacefrog

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Posted 05 February 2016 - 01:28 PM

I was looking at the growth in the one o'clock position on the edge of the last plate. If it was a contaminant of some sort it may compromise future transfers.


Good eye, Needles I didn't even see that.

It's so small and on the very edge though, you should easily be able to transfer from that plate if you are very clean and not pushing the scapel or other tool into the agar to cool the tip.

#10 invisibilitysyndrome

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Posted 24 February 2016 - 01:46 PM

First 2 pictures are TLY the pic of the 5 plates are AA+ and the rest are the GT

20160224_110639.jpg 20160224_110635.jpg 20160224_110539.jpg 20160224_110511.jpg 20160224_110506.jpg 20160224_110503.jpg 20160224_110459.jpg 20160224_110455.jpg

So here are a few more plates. I poured some more agar today to do more transfers.
I did take one gt plate and make some wbs jars to get a jump start on the fruiting process.

I look forward to growing these out. Especially the tly and aa+
Got my slant tubes already so im ready for some long term storage.
Tired of always going back to spore syringe if i run into spawn issues.

Edited by invisibilitysyndrome, 24 February 2016 - 07:48 PM.

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#11 peacefrog

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Posted 24 February 2016 - 05:42 PM

Looking real good, my friend.

My only advice would be to transfer smaller sections of agar. The smaller the sections you isolate, the better chance you have of isolating a mono culture in the fastest time:
image.jpeg image.jpeg

Nice looking plates!

Edited by peacefrog, 24 February 2016 - 05:48 PM.

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#12 invisibilitysyndrome

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Posted 24 February 2016 - 07:46 PM

@peacefrog right on makes sense the smaller the section the more precise the selection...
thnx for looking in on my plates.
The only myco books i have are as PDF format and they are difficult for me to navigate so i get more useful information from this site. Basicly im going blind based only on what ive seen and read on here so i do appreciate all the insight...

#13 whitethumb

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Posted 25 February 2016 - 05:12 AM

nicely done. your plates look good. pretty much what everyone said is spot on. you said your aa+ is from a clone? either way nice, i'll be watching.
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#14 invisibilitysyndrome

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Posted 25 February 2016 - 10:24 AM

nicely done. your plates look good. pretty much what everyone said is spot on. you said your aa+ is from a clone? either way nice, i'll be watching.


The GT is a tissue clone the plates in the photos are the 3rd transfer of GT. Gonna do 15 more plates today try and clean up the aa+ and tly few more times and hopefully get a true isolate from the GT.
Thnx fer stopping by..
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#15 invisibilitysyndrome

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Posted 25 February 2016 - 01:38 PM

20160225_125539.jpg 20160225_125533.jpg 20160225_125528.jpg 20160225_125524.jpg 20160225_125519.jpg

GT first 2
TLY 2nd 2
AA+ 3rd set of photos

GT might now be with a little luck a pure culture. We shall see
The others are in the beginning of transfers but you can see in the photos where the wedges were taken from.
Well on my way to some, fingers crossed , good fruiting cultures.

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#16 Needles

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Posted 25 February 2016 - 11:49 PM

GT first 2
TLY 2nd 2
AA+ 3rd set of photos
GT might now be with a little luck a pure culture. We shall see
The others are in the beginning of transfers but you can see in the photos where the wedges were taken from.
Well on my way to some, fingers crossed , good fruiting cultures.

Your plates look awesome, love that rizmorphic mycelium. Thoes jars of grain look real nice for what I could see too. Your definitely well on your way to the results you are looking for. Nice work getting your strain isolations......
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#17 invisibilitysyndrome

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Posted 26 February 2016 - 10:43 AM

GT first 2
TLY 2nd 2
AA+ 3rd set of photos
GT might now be with a little luck a pure culture. We shall see
The others are in the beginning of transfers but you can see in the photos where the wedges were taken from.
Well on my way to some, fingers crossed , good fruiting cultures.

Your plates look awesome, love that rizmorphic mycelium. Thoes jars of grain look real nice for what I could see too. Your definitely well on your way to the results you are looking for. Nice work getting your strain isolations......
Right on thnx man ya the jars are
A Gt AA+ and Falbino MS injected had trouble getting the falbino to take hold on the agar so i tried the grain hopefully i can get a good plate from a grain sample. Been so long since ive done MS i thought something was wrong w the jars.
Didnt see growth for 6 7 days.
Now i remember why being new to this all can be nerve wracking. Ive forgotten how long it can take MS vs a g2g etc...

#18 invisibilitysyndrome

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Posted 27 February 2016 - 07:15 PM

20160227_185847.jpg 20160227_185746.jpg

Little update
Plates are the latest transfers. They show good growth for only a couple days.

The jars are of the agar to grain done on the 24th.
The agar was one of the nice GT plates i had and decided to multiply it and will ultimatly try and grow em out. ...
Updates to come as they are available

Id like to amend the first post i made in this thread to clairify things.
The first 6 plates are TLY w the first 2 being the original MS and numbers 3 4 5 6 being the plates made from the first 2.
The next 4 are the AA+ that are of 2nd transfers
The last plate is of a GT that was at transfer #3.
I had several plates of GT that looked like the last but didnt show them.
one of those plates was used to make the jars in this post.
I hope ive not confused things. Id like to keep this thread open and on going so a little bit of clairity goes a long way....cheers

Edited by invisibilitysyndrome, 27 February 2016 - 07:25 PM.


#19 invisibilitysyndrome

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Posted 29 February 2016 - 12:29 PM

Looking real good, my friend.
My only advice would be to transfer smaller sections of agar. The smaller the sections you isolate, the better chance you have of isolating a mono culture in the fastest time:
attachicon.gifimage.jpegattachicon.gifimage.jpeg
Nice looking plates!


@peacefrog i just realized looking at the photo you showed here and in your other threads.
You put 2 pieces of agar on a plate/jar...
Is this just to maximise surface are?
Do you not feel it beneficial to give the myc more room to grow out for selection? Dosnt a small growth pattern make it harder to identify traits vs a full plate ?

#20 invisibilitysyndrome

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Posted 01 March 2016 - 12:46 PM

Some more pictures of the plates.
The number in the photo is the transfer those particular plates are on.
Photos taken late on the 29th feb.
20160229_203954.jpg 20160229_203657.jpg 20160229_204108.jpg




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