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OfflineTeaSippinHippie
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Registered: 11/09/13
Posts: 621
Loc: small sticks town
Last seen: 3 years, 2 months
Not another Agar post.
    #23792049 - 11/01/16 04:47 PM (7 years, 2 months ago)

Now that I'm in my own home I've been gathering things for agar work,i have the following:

- 1, 120qt SAB.
- 1, 500ml Borosilicate Glass bottle with
  Gl45 lid.
- multispore syringe.
- stainless steel scalpel with 20 #11 
  blades.
- PC
- dry MEA mix.
- 6 Borosillicate glass petris.

I've been doing my reading but feel im missing some information, been following the recent agar pages but just don't feel confident.

Before i begin the log i have a few questions.

-What items will i need in the SAB during.

-What do i wrap the petris in while PCing?
  I read i can put them in a small plastic container, doesn't plastic melt in the PC.

-how many dishes/transfers will it take for a pretty close isolated strain from MS?

Thank you all in advanced i wish i could've done this years ago.

I'm also about able to start donating to the shroomery :aweohyou: love this place.


--------------------
I LOVE THE SHROOMERY, GOODVIBES TO EVERYONE.
MudaFuka said:
There's nothing wrong with making mistakes as long as you don't make the same ones twice.


Edited by TeaSippinHippie (11/01/16 07:02 PM)


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Offlinespacechildo
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Re: Not another Agar post. [Re: TeaSippinHippie]
    #23792081 - 11/01/16 04:58 PM (7 years, 2 months ago)



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OfflineTeaSippinHippie
Male


Registered: 11/09/13
Posts: 621
Loc: small sticks town
Last seen: 3 years, 2 months
Re: Not another Agar post. [Re: spacechildo]
    #23792161 - 11/01/16 05:24 PM (7 years, 2 months ago)

I've read up on all that, i got my basic plan down for preparing the agar, I've been reading a tek by bodhisatta and one by stro.

I just need a few questions answered and guidance along the way till i get a successful grow using agar, but any comments are welcome, i am saving up for more dishes, that kind of glass is expensive though haha


--------------------
I LOVE THE SHROOMERY, GOODVIBES TO EVERYONE.
MudaFuka said:
There's nothing wrong with making mistakes as long as you don't make the same ones twice.


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Offlinespacechildo
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Re: Not another Agar post. [Re: TeaSippinHippie]
    #23792284 - 11/01/16 05:54 PM (7 years, 2 months ago)

why buy glass if you're doing pour agar?


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OfflineTeaSippinHippie
Male


Registered: 11/09/13
Posts: 621
Loc: small sticks town
Last seen: 3 years, 2 months
Re: Not another Agar post. [Re: spacechildo]
    #23792507 - 11/01/16 06:55 PM (7 years, 2 months ago)

I'll be trying both, and i like that i can re use them, and put them in the oven if i need to.


--------------------
I LOVE THE SHROOMERY, GOODVIBES TO EVERYONE.
MudaFuka said:
There's nothing wrong with making mistakes as long as you don't make the same ones twice.


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Invisibledankington
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Re: Not another Agar post. [Re: TeaSippinHippie]
    #23792566 - 11/01/16 07:11 PM (7 years, 2 months ago)

you'll soon grow very tired of working that way. When you start going through 20+ plates a week, you'll understand. If you're being environmentally friendly, do no-pour. Otherwise, do yourself a favor and buy a few hundred dishes.


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Offlinenumnum59
Pro-Am Mycologist


Registered: 12/07/11
Posts: 1,189
Last seen: 5 months, 22 days
Re: Not another Agar post. [Re: dankington]
    #23792624 - 11/01/16 07:23 PM (7 years, 2 months ago)

listen to these guys, i started agar maybe 2 weeks ago and im through 50 dishes so far. i used pastywhytes no pour tek for 25 of them and its awesome because they are reusable. im going to use them to isolate then xfer to a petri for storage.


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OfflineSpore Ninja
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Re: Not another Agar post. [Re: TeaSippinHippie]
    #23792666 - 11/01/16 07:30 PM (7 years, 2 months ago)

Quote:

TeaSippinHippie said:
Now that I'm in my own home I've been gathering things for agar work,i have the following:

- 1, 120qt SAB.
- 1, 500ml Borosilicate Glass bottle with
  Gl45 lid.
- multispore syringe.
- stainless steel scalpel with 20 #11 
  blades.
- PC
- dry MEA mix.
- 5 pyrex glass petris.

I've been doing my reading but feel im missing some information, been following the recent agar pages but just don't feel confident.

Before i begin the log i have a few questions.

-What items will i need in the SAB during.

-What do i wrap the petris in while PCing?
  I read i can put them in a small plastic container, doesn't plastic melt in the PC.

-how many dishes/transfers will it take for a pretty close isolated strain from MS?

Thank you all in advanced i wish i could've done this years ago.

I'm also about able to start donating to the shroomery :aweohyou: love this place.




Awww..another agar angels gets his wings. Always cool to see.  :goodluck:

Rule 1- don't overthink it- you just need to start pouring and inoculating plates. The more you do, the better you'll get.
Do some "practice" plates first if you want to, so there's no stress about contams or possibly losing your work to unforeseen events.
Heck, you can try pouring some plates in the open air if you just want to practice. Then just re-sterilize if glass, or toss them if they're plastic. (now you can start to see whey disposable plastic makes more sense)
So practice all the steps- cooking agar AND pouring plates til you're comfortable with it. This will answer your 1st question for you.

2- I'd really, really suggest using disposable, pre-sterilized plates. 
You might need 10 or more plates at one time.
Buy 3,4 or 5 sleeves of 100mm plates from FP or a vendor.

If you want to use glass- either buy a rack for them, or wrap them in foil. As far as I know- it's impossible to PC a petri dish with agar in it.
Look at the plastic container's recycling code- if has a "5" inside the triangle, it will survive the PC. I know a 5 (polypropylene or PP) will survive a PC, I don't think any of the other plastics will.

Here's how RR described the isolation process(I'm paraphrasing):
This assumes a clean culture with no contam issues-

Make a small line/wipe of your spores on agar.
AS SOON as the myc appears- cut small sections the size of a grain of rice and transfer them to separate dishes.

Then, let those grow out until you can see some sectoring.

Choose sectors from each dish, and cut small samples to transfer to new dishes.

Choose healthy, vigorous myc from the last transfers to transfer to grain.

Make mini casings or mini tubs to fruit out the grain so you can see which isolate yields the best.


So that means you need 1-3 dishes for your initial swipe(s), then in a couple days, you'll need 5-10 dishes to accept the 1st tiny samples.
Then you'll probably need 10+ dishes to accept the samples after you see sectoring.
You'll need at least 10-15 dishes.

Have you tried mixing some agar and pouring plates yet?? Let's break those babies in!!


--------------------
I'm interested in Dr. Pollock's work and his (especially older) strains.
Anyone have pix of his Shroommobile RV??

PSA: Protonmail offers free, encrypted email hosted in privacy loving Switzerland.
Red Phone / Signal offer free encrypted phone calls and texting between users.

All of these programs are available free for PC and Android or apple and are available in the respective stores or online by googling the names.
Stay safe out there!

The Electronic Freedom Foundation's review of text and messaging program's security is here>>> https://www.eff.org/node/82654


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OfflineTeaSippinHippie
Male


Registered: 11/09/13
Posts: 621
Loc: small sticks town
Last seen: 3 years, 2 months
Re: Not another Agar post. [Re: numnum59]
    #23792667 - 11/01/16 07:30 PM (7 years, 2 months ago)

I won't be doing agar a lot, so after i buy a few more sets of 6 ill be ok. Can any of you answer my questions up above?


--------------------
I LOVE THE SHROOMERY, GOODVIBES TO EVERYONE.
MudaFuka said:
There's nothing wrong with making mistakes as long as you don't make the same ones twice.


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OfflineTeaSippinHippie
Male


Registered: 11/09/13
Posts: 621
Loc: small sticks town
Last seen: 3 years, 2 months
Re: Not another Agar post. [Re: TeaSippinHippie]
    #23792700 - 11/01/16 07:37 PM (7 years, 2 months ago)

Thank you ninja for answering my questions, if i have too many contaminated dishes at first I'll buy some pre sterilized plastic ones and put the glass up till im better and can go through it with less dishes.


--------------------
I LOVE THE SHROOMERY, GOODVIBES TO EVERYONE.
MudaFuka said:
There's nothing wrong with making mistakes as long as you don't make the same ones twice.


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Offlinenumnum59
Pro-Am Mycologist


Registered: 12/07/11
Posts: 1,189
Last seen: 5 months, 22 days
Re: Not another Agar post. [Re: Spore Ninja]
    #23792705 - 11/01/16 07:38 PM (7 years, 2 months ago)

Before i begin the log i have a few questions.

-What items will i need in the SAB during.

-What do i wrap the petris in while PCing?
  I read i can put them in a small plastic container, doesn't plastic melt in the PC.

-how many dishes/transfers will it take for a pretty close isolated strain from MS?


in the sab i usually just have my clean dishes and my dishes im xfering. if im pouring ill have my petri dishes and the bottle with the agar.

not sure about what you wrap them in while pcing, if theyre glass then just put them on a rack or possibly wrap them in tin foil.

it can vary, i have done 2 xfers on several strains so far and im not close to getting an isolate.


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Invisibledankington
The Stranger
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Registered: 03/14/15
Posts: 4,577
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Re: Not another Agar post. [Re: TeaSippinHippie]
    #23792710 - 11/01/16 07:39 PM (7 years, 2 months ago)

You'll have a lot of contams. Could you imagine if I used glass petris? This stack is all garbage to me, and from about 1 week of work :frown:



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OfflineSpore Ninja
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Registered: 10/25/16
Posts: 161
Last seen: 5 years, 6 months
Re: Not another Agar post. [Re: TeaSippinHippie]
    #23792794 - 11/01/16 07:59 PM (7 years, 2 months ago)

Quote:

TeaSippinHippie said:
I won't be doing agar a lot, so after i buy a few more sets of 6 ill be ok. Can any of you answer my questions up above?




Hope I just did.

You also need:
a couple bottles of 70% alcohol
latex or nitrile gloves
paper towels
Oust, or Ozium
maybe some Clorox wipes
an alcohol lamp or good lighter/torch
an inoculating loop (or sterile swabs. You can DIY a good loop - I heard you can use toaster heating elements since they're made from nichrome)

All of that is available from Walmart. You can get a cheap toaster at a thrift store. I scored a set of 7 small plastic bins at Goodwill today that are perfect for mini casings.

DO NOT USE FLAMMABLE ALCOHOL INSIDE A GLOVEBOX WITH FLAME OR A LIGHTER!
(Ask me how I know)


Edited by Spore Ninja (11/01/16 08:17 PM)


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OfflineTeaSippinHippie
Male


Registered: 11/09/13
Posts: 621
Loc: small sticks town
Last seen: 3 years, 2 months
Re: Not another Agar post. [Re: Spore Ninja]
    #23792889 - 11/01/16 08:24 PM (7 years, 2 months ago)

I have most of that already from knocking up spawn bags. Should i keep any cleaning supplies inside the SAB? Like lysol?

And I'm pretty ocd when it comes to being clean before i work, i take shower get out go into work room, spray generous amount of lysol, put clothing on straight out of sterilized cycle on my washer n dryer, wipe area with 99.9 rubbing alcohol, then all tools and anything ill be touching, then my hands, gloves on, hands again then arms, then a hexagon mask.

And thats normally all without the SAB i normally don't use one for just inoculations, but for it ill add extra steps like cleaning the inside of the SAB and placing it on a soapy samp towel.


--------------------
I LOVE THE SHROOMERY, GOODVIBES TO EVERYONE.
MudaFuka said:
There's nothing wrong with making mistakes as long as you don't make the same ones twice.


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OfflineTeaSippinHippie
Male


Registered: 11/09/13
Posts: 621
Loc: small sticks town
Last seen: 3 years, 2 months
Re: Not another Agar post. [Re: dankington]
    #23792953 - 11/01/16 08:44 PM (7 years, 2 months ago)

20 pre sterilized dishes on the way :thumbup:


--------------------
I LOVE THE SHROOMERY, GOODVIBES TO EVERYONE.
MudaFuka said:
There's nothing wrong with making mistakes as long as you don't make the same ones twice.


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OfflineSpore Ninja
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Registered: 10/25/16
Posts: 161
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Re: Not another Agar post. [Re: TeaSippinHippie]
    #23793215 - 11/01/16 10:02 PM (7 years, 2 months ago)

Quote:

TeaSippinHippie said:
I have most of that already from knocking up spawn bags. Should i keep any cleaning supplies inside the SAB? Like lysol?

And I'm pretty ocd when it comes to being clean before i work, i take shower get out go into work room, spray generous amount of lysol, put clothing on straight out of sterilized cycle on my washer n dryer, wipe area with 99.9 rubbing alcohol, then all tools and anything ill be touching, then my hands, gloves on, hands again then arms, then a hexagon mask.

And thats normally all without the SAB i normally don't use one for just inoculations, but for it ill add extra steps like cleaning the inside of the SAB and placing it on a soapy samp towel.




Sometimes I'll keep my supplies in my glovebox if I don't have a dedicated room, or anywhere else to stash them.
Sounds like you have things covered pretty well.
There is a bit of an art to pouring agar- it's super messy if you spill it. You should practice pouring it (and cooking it)once or twice so you can see how to handle it.

You need to:
-lift the petri lid
-keep your fingers out of the way
-pour just the right amount of agar into the dish - too much and it spills or overflows and ruins that dish, too little and it might not work at all
-then give the bottle a slight twist as you finish each pour so it      doesn't drip. One misplaced drip can ruin a plate (or a stack!).
-then get the lid back on the peti
-wrap with saran wrap cut into a 2 inch strip, or wrap with Parafilm.
-then repeat it on the next petri in the stack. You work with the pertis in a stack, not dishes laid out individually.

Not really all that difficult, but it takes a bit of practice to get all that just right.
Oh, and you can't go too slow or the agar will harden up in the flask.:grin:


RR has videos showing pours, and there are several on Youtube that show pouring and SAB procedures.
"pouring agar plates"
"using SAB"


--------------------
I'm interested in Dr. Pollock's work and his (especially older) strains.
Anyone have pix of his Shroommobile RV??

PSA: Protonmail offers free, encrypted email hosted in privacy loving Switzerland.
Red Phone / Signal offer free encrypted phone calls and texting between users.

All of these programs are available free for PC and Android or apple and are available in the respective stores or online by googling the names.
Stay safe out there!

The Electronic Freedom Foundation's review of text and messaging program's security is here>>> https://www.eff.org/node/82654


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OfflineTeaSippinHippie
Male


Registered: 11/09/13
Posts: 621
Loc: small sticks town
Last seen: 3 years, 2 months
Re: Not another Agar post. [Re: Spore Ninja]
    #23793759 - 11/02/16 06:07 AM (7 years, 2 months ago)

The glass bottle i got too put the agar mix was recommended by all the big dogs on the site, gl45 lid prevents that unwanted drip.


--------------------
I LOVE THE SHROOMERY, GOODVIBES TO EVERYONE.
MudaFuka said:
There's nothing wrong with making mistakes as long as you don't make the same ones twice.


Extras: Filter Print Post Top
OfflineSpore Ninja
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Registered: 10/25/16
Posts: 161
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Re: Not another Agar post. [Re: TeaSippinHippie]
    #23794507 - 11/02/16 12:45 PM (7 years, 2 months ago)

Quote:

TeaSippinHippie said:
The glass bottle i got too put the agar mix was recommended by all the big dogs on the site, gl45 lid prevents that unwanted drip.




Sounds like your on a solid path and will do well...lol.. just maybe some 1st time jitters. Practice makes perfect. Don't treat the dishes like delicate little beauty queens, the want to get dirty and get used...often!

What's your first project going to be?


--------------------
I'm interested in Dr. Pollock's work and his (especially older) strains.
Anyone have pix of his Shroommobile RV??

PSA: Protonmail offers free, encrypted email hosted in privacy loving Switzerland.
Red Phone / Signal offer free encrypted phone calls and texting between users.

All of these programs are available free for PC and Android or apple and are available in the respective stores or online by googling the names.
Stay safe out there!

The Electronic Freedom Foundation's review of text and messaging program's security is here>>> https://www.eff.org/node/82654


Edited by Spore Ninja (11/02/16 12:51 PM)


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OfflineTeaSippinHippie
Male


Registered: 11/09/13
Posts: 621
Loc: small sticks town
Last seen: 3 years, 2 months
Re: Not another Agar post. [Re: Spore Ninja]
    #23794585 - 11/02/16 01:28 PM (7 years, 2 months ago)

I just want to mess around at first with MS on the dishes while my spawn bags finish colonizing, get my technique down and watch mycelium grow, get pointers and tips on here.
i might save any good dishes.

Once my bags are done and i spawn them and get a nice specimen i want to clone it.

Im not super nervous, i just wanted some questions answered before i started the fun.


--------------------
I LOVE THE SHROOMERY, GOODVIBES TO EVERYONE.
MudaFuka said:
There's nothing wrong with making mistakes as long as you don't make the same ones twice.


Extras: Filter Print Post Top
Offlinenumnum59
Pro-Am Mycologist


Registered: 12/07/11
Posts: 1,189
Last seen: 5 months, 22 days
Re: Not another Agar post. [Re: TeaSippinHippie]
    #23794619 - 11/02/16 01:39 PM (7 years, 2 months ago)

a couple tips i missed my first time that i wish i would have known.

bodhisatta -after you boil your agar and let the PC cool enough to pull it out "put them in 47ºC water bath. your agar jars are hotter than 47 so the water bath will warm up. once it cools back down to 47ºC then you're ready to pour." this will eliminate condensation on the lids which isnt necessarily a vector but it is annoying trying to see your growths through condensation.

i use a 750ml burtons gin bottle (~$7) for my agar, clean off the label and its long neck is perfect for pouring(any 750ml liquor bottle will work)

i also use brand new ziplock sandwich bags to seal the petri dishes due to the fact seran wrap is a bitch to cut how people say to do it for agar. it works so far for me.

if i think of any more, ill edit the post.


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