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tahoe
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Registered: 11/26/03
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Transfers, Isolates, and Sectors?? Please explain 1
#7745991 - 12/11/07 05:12 PM (16 years, 1 month ago) |
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The one thing that I am still unclear of when it comes to mycology is the difference between Isolates and transfers. I know what the definitions are but when does a transfer become an isolate?
I am assuming if one was to clone a mushroom to agar then this would already be an isolate of a strain and that you can not get away from these genetics.
If I were to start with spores on a petri and transfered a section of good looking mycelium from this petri to a fresh one; does this become an isolate?
I assume an isolate will not sector anymore but what is a sector? I thought sectors were areas of different growth.
How far away from the original spore laden petri do you folk like to get before you start selecting strains/isolates for cultivation.
Am I just way off?
-------------------- Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.
My Legacy https://www.shroomery.org/forums/showflat.php/Number/22140987#22140987 Teh=The I need to proofread
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RogerRabbit
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: tahoe] 1
#7746074 - 12/11/07 05:30 PM (16 years, 1 month ago) |
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A sector is a strain. A sector develops when compatible hyphae join to become a single organism, sharing genetics. Strains can also join with other strains via dikaryotic pairings, known as anastomosis. An isolate is a single strain, meaning you have transfered all the sectors to their own plates.
By taking tiny pieces of mycelium when I make transfers, I can often get to single sector isolates with three to four transfers. The dish in the picture on the left below is the result of two or three transfers. You can clearly see each sector by holding the dish up to the light. Tiny cuts were made in each of these sectors to move that mycelium to new dishes, and each of those became a single sector isolate. The right picture shows a dish that is still sectoring, and on the right is an isolated strain. Note there is no sectoring at all. Each and every isolate should be grown through the fruiting stage to determine the best performers. Prior to doing so, a small amount of mycelium should be carefully labeled and placed into the refrigerator on a petri dish or culture slant. This way, after fruiting, you can go back to an earlier stage in cell division for each future grow, capturing an aggressive strain. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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tahoe
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: RogerRabbit]
#7746160 - 12/11/07 05:49 PM (16 years, 1 month ago) |
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okay that is what I thought. Now for an outdoor species like ps cyanescens I do not want to do all this because that chance of getting a non fruiting sector which I would not know until next fall.
So as long as I do not get more then 2-3 transfers away from the original spore laden petri there will still be sectoring going on but this will be done while its colonizing the wood chips and the strongest fastest mycelium will win.
So both of these look pretty good one being 2 transfers away from the original and the other being 3. Well of corse minus the dark brown spot on the cyan dish. These were left over from last year and I just found them hiding in my closet

-------------------- Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.
My Legacy https://www.shroomery.org/forums/showflat.php/Number/22140987#22140987 Teh=The I need to proofread
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Deej3987
Deej3987

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Re: Transfers, Isolates, and Sectors?? Please explain [Re: tahoe]
#11828808 - 01/14/10 02:28 PM (14 years, 17 days ago) |
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Need some help i dont know if im sectoring this right, Im using (MEA) in the 1/2 pint jars or w/e size is the smallest and the strain is psilocybin Pink Buffalo.. Im refridgerating the sectors now but i dont think im doing it right, or maybe i am i just need some guidence if anyone can help me

^ This is the first sector.. If you look you can see where i used the inoculate loop to sector onto the next one..

^ Sector 2...

^ Sector 3...

^ Sector 4... and this is where ive stopped because the myc isnt looking as rhizimorphic as it did in the very first sector..
Can anyone help me with this??? I need some feedback please, everything on here about sectoring doesn't give me good enough pointers so i dont know if im doing it right.. ive seen all the pics and read up on it but sector 4 doesn't look as rhizimorphic as the other sectors.. maybe its just me idk if i have it or not please if someone can help me out a little more i would really appriciate it thanx!!
-------------------- My Trade List Printing All info I post about myself, what I am doing, and what I have done is fake; and therefore, holds no truth and can not be used to prove any previous action of mine.
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Necco
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: Deej3987]
#11835175 - 01/15/10 01:23 PM (14 years, 16 days ago) |
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You should have started a new thread instead of restarting one that is years old (since it isn't directly related). Also, I'm pretty sure your post would be better suited to non-advanced Mushroom Cultivation.
BTW, your spelling & grammar could use some help. In English, "I" is capitalized and "I'm" has an apostrophe. Refrigerator is not spelled refridgerator, etc. Jesus, I could go on and on.
You don't have psilocybin pink buffalo, you have Psilocybe cubensis var. "Pink Buffalo." Psilocybin is a psychoactive chemical inside the mushroom.
What you call "sector" in your post is actually "transfers." For instance, there are countless sectors in your transfer 1 (labeled sector 1), dozens of sectors in your transfer 2 (labelled sector 2). It looks to me like you still have many sectors in transfer 4.
You say "everything on here about sectoring doesn't give me good enough pointers" when it is obvious you haven't read everything on here on the topic.
Here is a video that can get you started, although it would really be helpful if you could watch the entire video, either way it is a good start: http://www.mushroomvideos.com/Strain-Isolation
Here is a little tutorial along the same lines: Strain Isolation on Agar. w/pix: http://archives.mycotopia.net/discus/messages/5/41777.html?1067230771
This is some good info too: http://www.shroomery.org/forums/showflat.php/Number/545485#545485
I'm sure there is tons more if you look, since I found that all in about 2 minutes with a couple searches.
Your question "can somebody help me with this?" is vague, help you with what? Next time try to post a question that is very specific if you want any good answers. One thing I can suggest is that you keep more of your transfers. You should have done something more like this: 3 transfers from your first dish, make 3 transfers off of those dishes (now you have 9 plates) and make two transfers off of those plates (now you have 18) and then select your best looking sectors for further transfering; in the end you would keep many isolates for testing. It looks like you only kept one plate on your 4th transfer, you should have over a dozen by then so you can compare the different isolations/colonies.
None of this post was meant to make you feel bad, I'm just trying to give you some good advice.
--------------------
"Now ether was substituted for chloroform, and the difference of their phenomena noted, and now some other exhilarant, in the form of an opiate or stimulant, was the instrument of my experiments, until I had run through the whole gamut of queer agents within my reach..." I can do everything!!
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teesionbear


Registered: 12/13/09
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: Necco]
#11845242 - 01/17/10 02:14 AM (14 years, 14 days ago) |
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Quote:
mubba said: .
Here is a video that can get you started, although it would really be helpful if you could watch the entire video, either way it is a good start: http://www.mushroomvideos.com/Strain-Isolation
Thanks for that link man! Some good tips in there about speeding up isolation.
-------------------- MY TRADE LIST
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Deej3987
Deej3987

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Re: Transfers, Isolates, and Sectors?? Please explain [Re: teesionbear]
#11846554 - 01/17/10 11:26 AM (14 years, 14 days ago) |
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Thanks for the criticism, I really do appreciate it. I thought this thread would be good for what I was doing. Ill post more on my progress once I get those petri dishes I ordered and Ill be sure to make more transfers next time.
I have read a lot about strain isolation on here, but I couldn't find dummy proof picture tutorials/demonstrations on where the different sectors are exactly. It doesn't look like I'm doing it correctly. My myc isn't as stringy (Rhizomorphic) as it appeared in the 2 o'clock or 5 o'clock sectors on the first *transfer* <not sector) on the left.
Thanks again for the links. I have seen the "Lets Grow Some Mushrooms" video a few times (not the full version). I'm looking for more or better details on where I should sector the myc...
I guess my question was where do I need to sector to get the best rhizomorphic mycilium?
-------------------- My Trade List Printing All info I post about myself, what I am doing, and what I have done is fake; and therefore, holds no truth and can not be used to prove any previous action of mine.
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PrimalSoup
hyperspatial illuminations



Registered: 11/17/09
Posts: 13,568
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: Deej3987]
#11849775 - 01/17/10 09:27 PM (14 years, 14 days ago) |
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Quote:
I guess my question was where do I need to sector to get the best rhizomorphic mycilium?
That's kind of a toughie - it's why you're being advised to do more transfers off each plate and grow them, then repeat, until you have a lot of examples to select from.
With some experience it's easier to judge which areas you want to transfer from, without that it's more of a guess. Doesn't mean you can't get lucky, but you won't know if there were other possibilities there on the plate that you missed. 
It might be helpful to use actual petri dishes instead of half-pint jars (?) as they're easier to work with and allow more space for growth. You can use glass ones or get the presterilized plastic ones easily enough. Advantage is you can grow the cultures out longer in between transfers, allowing you to judge how fast they're growing and, uhm, "rhyzomorphicity" since they change characteristics as they grow.
I'd also suggest you use a scalpel/knife not an innoculating loop for the transfers, allows you to pick out just the part you want (you don't need much), but this doesn't have to be fancy, just a tiny blade will work.
Yeah, the 4th transfer still is sectoring a lot, it doesn't have a lot of room there to stretch out, so to speak. But keep at it, it's fun to do and experience helps it make a lot more sense. 
Peace -PS
--------------------
if you stand too close to the machine it'll start to eat youPrimal's simple tested teks and projects: Wheat Prep 2.0 Acidic Tea Tek Potency Project!
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Necco
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: Deej3987]
#11850592 - 01/17/10 11:59 PM (14 years, 14 days ago) |
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Your #4 transfer looks like it has a good amount of healthy mycelium even though you may not see rhizo., so I wouldn't be too worried. Get some more isolates off of that one and you will may have some nice ones.
People say that nutrient dense agar can suppress rhizomorphic mycelium in some circumstances. One way you could promote the visualization of rhizomorphicity (that may not be a real word) is by using diluted agar. Figure out what dilution of agar mix you can use without problems; if you are using a pre-mixed formula then you should cut it back as far as you can without loosing the solidity of the agar, if you mix it yourself then cut back on the sugars and nutrients but leave the agar agar the same. When less nutrients are available, it causes the mycelium to search out nutrients elsewhere with its rhizo'.
I was also doing some isolation in jars and it was torture. Especially trying to look through the sides when it was growing. I got some real dishes and it made a world of difference. A lot of people prefer the plastic ones, but the glass ones are just as good if you ask me. They can be oven sterilized and reused.
--------------------
"Now ether was substituted for chloroform, and the difference of their phenomena noted, and now some other exhilarant, in the form of an opiate or stimulant, was the instrument of my experiments, until I had run through the whole gamut of queer agents within my reach..." I can do everything!!
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Deej3987
Deej3987

Registered: 12/08/09
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: Necco]
#11853099 - 01/18/10 01:26 PM (14 years, 13 days ago) |
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Thanks again you guys for the advice, Ill try using a scalpel for my next transfers and Ill wait until I have the petri dishes. Eventually Ill get the glass ones so I can reuse them I really like that idea, I don't want to keep buying them since Ill be using them a lot and Ill be sure to make more sectors from my 4th transfer.. I made my (MEA) and I'm going to need more soon so Ill try making it with less Malt Extract just to see what happens. Lots of good ideas to try out.
The myc looks different in the center than on the outer edge on the 4th transfer. So should I take sectors from the middle or just the outer edge?
Also where is a good place to get glass petri dishes? The ones I ordered are from sporeworks and idk if they are plastic or glass..
-------------------- My Trade List Printing All info I post about myself, what I am doing, and what I have done is fake; and therefore, holds no truth and can not be used to prove any previous action of mine.
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Noobey
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: RogerRabbit]
#11855453 - 01/18/10 07:24 PM (14 years, 13 days ago) |
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Quote:
RogerRabbit said: A sector is a strain. A sector develops when compatible hyphae join to become a single organism, sharing genetics. Strains can also join with other strains via dikaryotic pairings, known as anastomosis. An isolate is a single strain, meaning you have transfered all the sectors to their own plates.
By taking tiny pieces of mycelium when I make transfers, I can often get to single sector isolates with three to four transfers. The dish in the picture on the left below is the result of two or three transfers. You can clearly see each sector by holding the dish up to the light. Tiny cuts were made in each of these sectors to move that mycelium to new dishes, and each of those became a single sector isolate. The right picture shows a dish that is still sectoring, and on the right is an isolated strain. Note there is no sectoring at all. Each and every isolate should be grown through the fruiting stage to determine the best performers. Prior to doing so, a small amount of mycelium should be carefully labeled and placed into the refrigerator on a petri dish or culture slant. This way, after fruiting, you can go back to an earlier stage in cell division for each future grow, capturing an aggressive strain. RR

Not to thread jack but that cleared up a whole lot for me. Thanks RR
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PrimalSoup
hyperspatial illuminations



Registered: 11/17/09
Posts: 13,568
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: Noobey]
#11856250 - 01/18/10 09:23 PM (14 years, 13 days ago) |
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Quote:
The myc looks different in the center than on the outer edge on the 4th transfer. So should I take sectors from the middle or just the outer edge?
Outer edge IME, you're looking to maximize the growth differences and get it from the place where it's actively growing and dividing.
Quote:
Also where is a good place to get glass petri dishes? The ones I ordered are from sporeworks and idk if they are plastic or glass..
I just got another dozen off eBay to supplement the ones I've had almost forever.
Check regularly, they come up but don't stay long - the ones I got were Kimax and cost $23+shipping, brand new in old stock boxes - less than I paid years ago. If they don't say the material or they're cheaper they are plastic, and though plastic works fine, when you run out, you're out... 
Peace -PS
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if you stand too close to the machine it'll start to eat youPrimal's simple tested teks and projects: Wheat Prep 2.0 Acidic Tea Tek Potency Project!
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Necco
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: PrimalSoup]
#11856457 - 01/18/10 09:50 PM (14 years, 13 days ago) |
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Cynmar is a shroomery favorite for dishes. They have soda glass petris for quite a fair price (they also have the disposable petris as well as all kinds of other goodies).
http://www.cynmar.com/SearchByCategory.aspx?CategoryCode=271
If I could pull off getting a dozen Kimax petris for under $30 then I'd for sure go that route, but I don't see that kind of deal coming around too often.
--------------------
"Now ether was substituted for chloroform, and the difference of their phenomena noted, and now some other exhilarant, in the form of an opiate or stimulant, was the instrument of my experiments, until I had run through the whole gamut of queer agents within my reach..." I can do everything!!
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Deej3987
Deej3987

Registered: 12/08/09
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: Necco]
#12382727 - 04/13/10 12:24 AM (13 years, 9 months ago) |
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Hey sorry I haven't posted more on my progress with the agars I had to bail on all my work sadly, so next time I'll repost on a different thread starting from fresh.. buutt... Is there anywhere I can get huge ass petri dishes? that way the agars can cultivate more area. I'm talking about petri dishes 2 - 4 times bigger than the normal sized ones, maybe this would allow me to see potential sectors a lot easier?
-------------------- My Trade List Printing All info I post about myself, what I am doing, and what I have done is fake; and therefore, holds no truth and can not be used to prove any previous action of mine.
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fastfred
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: Deej3987]
#12384374 - 04/13/10 10:15 AM (13 years, 9 months ago) |
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You can get large square ones. The ones I have are about 8" x 8".
I don't know why you'd use them though, it's just going to waste a ton of media.
-FF
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Deej3987
Deej3987

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Re: Transfers, Isolates, and Sectors?? Please explain [Re: fastfred]
#12463346 - 04/27/10 01:20 AM (13 years, 9 months ago) |
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I want more space for the mycelium to colonize maybe sectoring would be easier?? I don't know, just something I want to try. Where did you say you got yours at?
-------------------- My Trade List Printing All info I post about myself, what I am doing, and what I have done is fake; and therefore, holds no truth and can not be used to prove any previous action of mine.
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fastfred
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: Deej3987]
#12464045 - 04/27/10 08:29 AM (13 years, 9 months ago) |
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I found mine at the recycling center actually. They make nice trays for printing, etc..
-FF
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RogerRabbit
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: Deej3987]
#12464955 - 04/27/10 11:47 AM (13 years, 9 months ago) |
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Quote:
Deej3987 said: I want more space for the mycelium to colonize maybe sectoring would be easier?? I don't know, just something I want to try. Where did you say you got yours at?
If the mycelium colonizes more, the various strains will tend to merge into a coherent whole, defeating the purpose of isolating. I use three section dishes, and never allow the mycelium to grow larger than a half-dollar. This way, individual strains can be isolated and tested. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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mainratte
humble
Registered: 03/23/10
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Re: Transfers, Isolates, and Sectors?? Please explain *DELETED* [Re: RogerRabbit]
#12476235 - 04/29/10 09:02 AM (13 years, 8 months ago) |
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Post deleted by mainratteReason for deletion: .
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PrimalSoup
hyperspatial illuminations



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Re: Transfers, Isolates, and Sectors?? Please explain [Re: mainratte]
#12477161 - 04/29/10 12:10 PM (13 years, 8 months ago) |
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Many will not fruit on agar. And the fruits on agar aren't necessarily going to help you differentiate unless you plan on growing them on agar. From my experience...
Peace -PS
--------------------
if you stand too close to the machine it'll start to eat youPrimal's simple tested teks and projects: Wheat Prep 2.0 Acidic Tea Tek Potency Project!
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RogerRabbit
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Re: Transfers, Isolates, and Sectors?? Please explain [Re: PrimalSoup]
#12479188 - 04/29/10 05:52 PM (13 years, 8 months ago) |
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Quote:
if i isolate some strains can i differenciate between them by simply fruiting them on the petri dishes
No, use brf cakes for quickest results. If you want the most accurate representation, fruit each isolate on the substrate you intend to use later. RR
-------------------- Download Let's Grow Mushrooms semper in excretia sumus solim profundum variat "I've never had a failed experiment. I've only discovered 10,000 methods which do not work." Thomas Edison
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