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OfflineEdmunter
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Agar questions and answers
    #23857076 - 11/22/16 11:44 AM (7 years, 2 months ago)

So I have some stuff going on with agar.  Id like to understand more about whats going on.

This P.Mex plate is DFA 8:10 did something strange on transfer.  It fluffed up b4 begining to row.  It happened to all 3 plates.  WTF is going on here then?

v

This P. Sandose has been producing spots and not growing like id like it to since the first transfer.  Do these have a history of this kinda growth



These PE plates both have gaps where it looks like it either hasnt liked the agar there or it's genetics?



This AA+ has produced a pin before the culture has reached the edge,  Im thinking I want those genetics as shes a white cracker bursting with life.  Transferring soon




India O plates are looking good.  Lots of nice sectors very clear



Thai Plate is looking good and getting closer to Isolate



Finally this Eringii clone from a shop bought mushroom is doing well and will be grown out to inoculate rye seed soon.



If you can help please add some comments.


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Re: Agar questions and answers [Re: Edmunter]
    #23857165 - 11/22/16 12:16 PM (7 years, 2 months ago)

If the agar is very nutritious the transfered pieces tend to fluff sometimes IME. You should not worry about that. I had this with different species that got nice growth later.
I have no experience with DF agar. Usually I use MEA.
The PE plate looks strange. Observe it and update this thread in 2 days.


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Re: Agar questions and answers [Re: enlightenment]
    #23857169 - 11/22/16 12:17 PM (7 years, 2 months ago)

:whathesaid:


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Re: Agar questions and answers [Re: bodhisatta]
    #23857549 - 11/22/16 02:38 PM (7 years, 2 months ago)

My last batch of PDA plates all look like those fluff balls you have there. Too much nutrients


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Re: Agar questions and answers [Re: Edmunter]
    #23859935 - 11/23/16 08:57 AM (7 years, 2 months ago)

I have 2 aa+ plates with pins.  After cloning  Is it ok to use these for LI and get decent results?


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Re: Agar questions and answers [Re: Edmunter]
    #23860028 - 11/23/16 09:24 AM (7 years, 2 months ago)

I would not spawn pins to bulk. Fruits are a vector for cubensis.


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Re: Agar questions and answers [Re: Munchauzen]
    #23860044 - 11/23/16 09:27 AM (7 years, 2 months ago)

So clone the pin and LI the plate so I get a  MS grow?  I kinda want some for xmas so im in a hurry.


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Re: Agar questions and answers [Re: Edmunter]
    #23860466 - 11/23/16 12:27 PM (7 years, 2 months ago)

Eat that pin merry x mas.


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Bag sealers are to bulky (my hood isn't that big)
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Re: Agar questions and answers [Re: Edmunter]
    #23862882 - 11/24/16 08:06 AM (7 years, 2 months ago)

These PE plate certainly dont like those parts of the agar.  Ok on 1 plate but to have 2 do the same suggests something else.  The other sides on both have nice sectoring going on.  I have take 3 transfers from 1 plate and hope for better all round genetics.




There she blows!  I was too late on one plate but the other was ok. so I made a LI and clones the Pin.





On a side note, can you use Cat food as a medium?


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Re: Agar questions and answers [Re: Edmunter]
    #23877196 - 11/29/16 03:53 AM (7 years, 2 months ago)

So the P.mex after the initial fluffing has grown out nice and cleanly and round.  Im thinking of using this for a master jar now.  Here are the latest photos



The lids a bit clouded due to old age but it looks fine to me.


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Re: Agar questions and answers [Re: Edmunter]
    #23877269 - 11/29/16 05:12 AM (7 years, 2 months ago)

I litterally just did the same thing a few days ago. My plate was super fluffy but perfectly round like yours. I cut it into 36 pieces and nocd up 6 jars with it.
The growth in jars is suprisingly the most rhizo I have ever had yet.




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:amanita2: Substrate surface conditions / Monotub prep and care :sporedrop:

Look around you... Everything you see exists inside the mind.
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Re: Agar questions and answers [Re: ComebackKid]
    #23877722 - 11/29/16 09:49 AM (7 years, 2 months ago)

One of the PE plates kinda colonised the patch.  What could of caused this?



I transferred away from the last plate but the new plate looks a bit sweaty.

The sectoring the on left of this plate is really clear and looking good.

Gunna transfer once more.


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Re: Agar questions and answers [Re: Edmunter]
    #23922686 - 12/13/16 01:27 PM (7 years, 1 month ago)

4th Transfer

PE



Made a LI with another similar which is going for it, so happy days.

Doubt I will be getting any Penis for christmas though which is a shame as I was really looking forward to some festive fun.

I heard somebody say they had never had a blob when they had straight cased which is interesting.

Where would you take from to get close to isolate?  I take it if I take one tiny wedge from the single stains that will isolate?  I know its not neccesarry but Id like a few isolates as ive never gone that far.



7 oclock looks quite uniform


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23983112 - 01/05/17 04:41 AM (7 years, 25 days ago)

So with my shinny silver pants im cooking the A again


So questions of the day
Are these aa+ pins?



What are these, blobs?


Is this a contamination?



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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23986062 - 01/06/17 09:08 AM (7 years, 24 days ago)

Quote:

Edmunter said:
So with my shinny silver pants im cooking the A again


So questions of the day
Are these aa+ pins?



What are these, blobs?


Is this a contamination?





The answer I got from another forum was

Yes
Knots
Yes

I m not sure if it is a contamination but there dont seem to be too many experts on this subject.



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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23986073 - 01/06/17 09:17 AM (7 years, 24 days ago)

:threadmonitor:


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23986086 - 01/06/17 09:25 AM (7 years, 24 days ago)

My next agar question is why does this culture do this?  It has done it since the first transfer away from spores and now I am at the 5th.





So when they start growing they leave an area like this.  Has anyone had this b4 and what causes it?  It has done it with every plate involved from the original plate.



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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23986127 - 01/06/17 09:54 AM (7 years, 24 days ago)

The dark stuff looks like sediment from the dog food...you're coloring your agar, correct?  If so, as the color fades, the sediment becomes more apparent...possibly.


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Re: Agar questions and answers(New questions) [Re: Mycolorado]
    #23986162 - 01/06/17 10:09 AM (7 years, 24 days ago)

that looks like a nice sector


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23986206 - 01/06/17 10:31 AM (7 years, 24 days ago)

Quote:

Edmunter said:
My next agar question is why does this culture do this?  It has done it since the first transfer away from spores and now I am at the 5th.





So when they start growing they leave an area like this.  Has anyone had this b4 and what causes it?  It has done it with every plate involved from the original plate.







Why does it grow round on itself and miss out that section?  I was thinking of using this to make a LI.  What do you think?


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23986227 - 01/06/17 10:39 AM (7 years, 24 days ago)

it will colonize that section if it isn't already colonized by a contam...

I dont find much use for LI and wouldn't make it unless I was absolutely certain I had a clean source.


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Re: Agar questions and answers(New questions) [Re: PsilocyBen17]
    #23986234 - 01/06/17 10:42 AM (7 years, 24 days ago)

But why would every plate from the spore plate in this chain do the same....Its bizarre.


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23986267 - 01/06/17 10:54 AM (7 years, 24 days ago)

its not bizzarre...myc grows from the outer edges....in a few days the plate will be colonized, unless bacteria/mold is preventing it from growing more, which is likely if this is the first transfer from spores.


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23986275 - 01/06/17 11:00 AM (7 years, 24 days ago)

Quote:

Edmunter said:
My next agar question is why does this culture do this? IT HAS DONE THIS WITH EVERY TRANSFER FROM THE SPORE PLATE AND NOW IM ON THE 5TH





So when they start growing they leave an area like this.  Has anyone had this b4 and what causes it?  It has done it with every plate involved from the original plate.






:getstoned:


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23986301 - 01/06/17 11:14 AM (7 years, 24 days ago)

Like ben pointed out, could very well be avoidance.  I've had growth like that on plates that are a couple transfers from spore syringe that were dirty initially...The plate of APE pins I posted earlier on pic of the day exhibited exactly what you're showing and the pins are secreting metabolites, which as we know, is sometimes indicative of contams.


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Re: Agar questions and answers(New questions) [Re: Mycolorado]
    #23986373 - 01/06/17 11:49 AM (7 years, 24 days ago)

So I really need to use a T or sandwich agar to be sure to not carry it for a 6th time?


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23986381 - 01/06/17 11:52 AM (7 years, 24 days ago)

If you look at above post there is a plate from this chain posted on the 29/11


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #23986789 - 01/06/17 03:09 PM (7 years, 24 days ago)

I'd transfer till it showed more uniformity and then run it, but that's just me. Overall, your culture looks like nice even clean growth...you could always take a transfer to continue to isolate and run a small tester with the rest...


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Re: Agar questions and answers(New questions) [Re: Mycolorado]
    #23986872 - 01/06/17 03:35 PM (7 years, 24 days ago)

Quote:

Mycolorado said:
I'd transfer till it showed more uniformity and then run it, but that's just me. Overall, your culture looks like nice even clean growth...you could always take a transfer to continue to isolate and run a small tester with the rest...




Good plan.


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #24071509 - 02/07/17 01:50 AM (6 years, 11 months ago)

So I read about Roger Rabbit making a master culture from a clone.  What is the definition of this and how can you continue on a culture unless it is a mono?  Im thinking unless you isolate it will have different vectors from different points and so not going to produce the same results.  For the sake of this thread can somebody either give me a comprehensive answer or link me to somewhere I can get my teeth into about it?


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #24071755 - 02/07/17 06:40 AM (6 years, 11 months ago)

Clones man... Clones are just that, clones... If you like a certain phenotype, size, clustering tendencies you take a clone from that... The grows from that clone will be pretty damn close to what you cloned from.


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Re: Agar questions and answers(New questions) [Re: LocN9ne]
    #24071759 - 02/07/17 06:42 AM (6 years, 11 months ago)

Idk if anyone answered your question about your p.Mex fluffing up like that... But that is just what that myc looks like, it doesn't get rhizo like cubensis does.


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #24071895 - 02/07/17 08:45 AM (6 years, 11 months ago)

Quote:

Edmunter said:
So I read about Roger Rabbit making a master culture from a clone.  What is the definition of this and how can you continue on a culture unless it is a mono?  Im thinking unless you isolate it will have different vectors from different points and so not going to produce the same results.  For the sake of this thread can somebody either give me a comprehensive answer or link me to somewhere I can get my teeth into about it?



You could squirt MS on a dish and take a wedge and make a slant. You can have a culture of a single strain or a culture that's 100s
The culture is what's slanted and saved. it's genetic makeup doesn't matter. Slanting is just a way to preserve genetics


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Re: Agar questions and answers(New questions) [Re: bodhisatta]
    #24072136 - 02/07/17 11:08 AM (6 years, 11 months ago)

Ok so Ive clone 6 plates from some PE pins 2nd transfer, so if I keep one of them as a master I can use the rest as I like and they will all be similar grows?


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #24072172 - 02/07/17 11:22 AM (6 years, 11 months ago)

Likely


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Re: Agar questions and answers(New questions) [Re: bodhisatta]
    #24072181 - 02/07/17 11:24 AM (6 years, 11 months ago)

Ok im getting my new pressure cooker soon and so will test them to see


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #24215390 - 04/03/17 01:39 PM (6 years, 9 months ago)

I took some in-vitro pins from some old AA+ forgotten plates and transferred to 4 new plates.  I didn't see any mold in the original, however I managed to have satellites in every plate in this batch.

Due to life getting in the way 12 hours passed from when I wanted to clean/transfer and when I did.  By that time the satellites had turned green,  So I just quickly transferred to 4 new plates.

My thinking is that the second I see growth from these to transfer to new plates to hopefully have something I can use.

My question is:  Green is nasty and in reality the next plate is going to be contaminated anyway so is it worth using your SAB, or just go open air and use the mycs speed to clean keeping is out of your sterile area?  I did it in my SAB and I hope I dont live to regret it.

Sound advise needed.


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #24215609 - 04/03/17 03:09 PM (6 years, 9 months ago)

Don't do it in open air ever.

Take a transfer as soon as it leaps off the original transfer try not to get any of the original transfer.

I just transferred some plates that were green as long as your sterile technique is on point it shouldn't be contaminated.

A Sab is exactly that still air the spores on a plate need air currents to spread. Or even a little bump.


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If I ever give out misinformation please inform me so I can have the correct information. :cheers:

Tmethyl said:
Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy.

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Re: Agar questions and answers(New questions) [Re: van hatton]
    #25646036 - 11/29/18 05:07 AM (5 years, 2 months ago)

So I just took a clone sample.  If the agar grows clean would you use this to inoc or transfer a few times?


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #25646038 - 11/29/18 05:09 AM (5 years, 2 months ago)

Few transfers at least 1 or 2


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If I ever give out misinformation please inform me so I can have the correct information. :cheers:

Tmethyl said:
Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy.

Caps McGee said:
:thumbsup:
Fun part is figuring out what works best for you


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Re: Agar questions and answers(New questions) [Re: van hatton]
    #25646047 - 11/29/18 05:19 AM (5 years, 2 months ago)

If I use 10:8 nute for speed would you advice using a higher nutes before transfer or doesnt it matter and reasons please.


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #25646054 - 11/29/18 05:26 AM (5 years, 2 months ago)

Uh.

Use a low ratio 2% is standard I use 1.5-1.75%

I like to keep my stuff constant so the mycelium doesn't make any strange moves on me and end up looking weird on me.

Stick with the ratio that works for u.


Reason I wouldn't do higher nutes specifically is because rizo mycelium doesn't happen very often on higher nutes plates. The nice thing about rizo is it's super easy to tell that from contams. So I shoot for rizo.

Does that make sense? :bongload:


--------------------
If I ever give out misinformation please inform me so I can have the correct information. :cheers:

Tmethyl said:
Chuck Norris once roundhouse kicked a monotub that wasn't pinning fast enough. The force of the kick rearranged the genetics of the mushrooms, we now call them Penis Envy.

Caps McGee said:
:thumbsup:
Fun part is figuring out what works best for you


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Re: Agar questions and answers(New questions) [Re: van hatton]
    #25646325 - 11/29/18 09:46 AM (5 years, 2 months ago)

One of the TCs was saying lower nute make nicer looking plate but its best to use higher nute plates to inocutate with.  Im just trying get confirmation here.


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #25646336 - 11/29/18 09:54 AM (5 years, 2 months ago)

I run 2% nutes across the board.


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Re: Agar questions and answers(New questions) [Re: Mycolorado]
    #25698443 - 12/23/18 02:34 PM (5 years, 1 month ago)

So if you wanted to start a new master how long should you leave a plate b4 using it.  I took a dime size from a plate the other day and cut just inside the outer growth.  My thinking is that this part is clean and even if the plate has something funky on it I should be ok.

What's your thinking/experience on this subject?


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #25699528 - 12/24/18 07:17 AM (5 years, 1 month ago)

Looks good so far


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #25727030 - 01/06/19 11:35 AM (5 years, 24 days ago)

Ive started some new videos to see how my technique is working.



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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #25727032 - 01/06/19 11:38 AM (5 years, 24 days ago)



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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #25727037 - 01/06/19 11:39 AM (5 years, 24 days ago)



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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #25737460 - 01/11/19 04:52 AM (5 years, 19 days ago)

So ive had a bit of a eureka moment that what im trying to do is grow a culture that is 100% clean on the culture itself regardless of whether the plate has a few spots.  In these photos I am quite happy they are clean and wondered what options I have?  Can you transfer out even when its really small to get rid of side contaminations if you are still learning your sterile technique?  Can you comment on these?


This looks clean

This looks clean

This looks clean

From a pin that appeared on a plate 1 transfer away from use



Edited by Edmunter (01/11/19 05:02 AM)


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Re: Agar questions and answers(New questions) [Re: Edmunter]
    #25737476 - 01/11/19 05:03 AM (5 years, 19 days ago)

From PE transfer




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