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Offlineeelrd72
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print to agar
    #18894307 - 09/26/13 02:32 PM (10 years, 4 months ago)

when using an inoculation loop on a print to agar, do you just scrape the print...or do you use a drop of water to lift some spores? Also How long should I expect before I see any growth on the plate?


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Invisible36fuckin5
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Registered: 08/11/03
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Re: print to agar [Re: eelrd72]
    #18894318 - 09/26/13 02:35 PM (10 years, 4 months ago)

Just rub it around a little bit. If you use water, you'll fuck up the rest of your print.

Cubensis can take up to a couple weeks to germinate. Most of the time you'll see something in a week or less


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


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InvisibleFooMan
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Re: print to agar [Re: eelrd72]
    #18894432 - 09/26/13 02:57 PM (10 years, 4 months ago)

Also, don't go crazy trying to scrape a ton of spores. They'll be on the loop even if you don't see them. If getting an isolate is your goal, the less spores you use the better. More spores means more sectoring and therefore more transfers before you get an isolate.


--------------------

Quick WBS Prep


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Offlineeelrd72
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Re: print to agar [Re: FooMan]
    #18894516 - 09/26/13 03:17 PM (10 years, 4 months ago)

thanks for the info...any idea how long it takes before I see anything? One more question..should i put upside down..because i have some humidity build up..from pouring too hot? and should i put in incubation chamber at 80 degree?


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Invisible36fuckin5
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Re: print to agar [Re: eelrd72]
    #18894523 - 09/26/13 03:19 PM (10 years, 4 months ago)

No incubation, and store them right side up.


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


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OfflinePantheist
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Registered: 06/05/16
Posts: 38
Last seen: 3 years, 4 months
Re: print to agar [Re: eelrd72]
    #23562050 - 08/21/16 03:00 AM (7 years, 5 months ago)

If working in a SAB I was concerned about flame sterilising the inoculation loop after wiping down all the insides with isopropyl alcohol! If anyone can clear up the procedure for me that would be great.

I thought it would be best to post in / bump this existing thread than to start a new one, so all the info is in one place. I did look through the forum but couldn't find an answer.


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Offlinemushpunx
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Re: print to agar [Re: Pantheist]
    #23562076 - 08/21/16 03:57 AM (7 years, 5 months ago)

]
If working in a SAB I was concerned about flame sterilising the inoculation loop after wiping down all the insides with isopropyl alcohol! If anyone can clear up the procedure for me that would be great.

I thought it would be best to post in / bump this existing thread than to start a new one, so all the info is in one place. I did look through the forum but couldn't find an answer.





There is no reason to wipe the insides with ISO.. its un nessacary and dangerous. If your SAB needs cleaning wash with dish soap and water.
You can't/dont need to anyways sterilize an SAB.

To prep a still air box you use a spray bottle of water with a little dish soap in it. You spray the walls lightly and the air a little and let it settle for a while. The soapy water will help trap airborne contams/settle the air quicker.

You do however, want to wipe all jars/plates down with ISO before placing in your SAB, and wash your gloves with ISO before and frequently during work.


To i nnoculate with a loop you would flame sterilize the loop untill red hot, cool it in the recieving blank agar plate, rub a tiny section of the print, and then swipe across the plate in a zig zag pattern.
When you cool the loop it gets some agar on it which helps to pick up spores off the platen


--------------------

Amateur Mycologists United
AMU Q&A


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Invisible404
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Re: print to agar [Re: mushpunx]
    #23562081 - 08/21/16 04:02 AM (7 years, 5 months ago)

i wipe the insides of my SAB every time with iso. I also use the torch lighter inside with no problem. I can't for safety reasons suggest or condone this, but in my experience i have never had a problem and i've been doing it this way for several years.


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Offlinemushpunx
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Re: print to agar [Re: 404]
    #23562130 - 08/21/16 05:04 AM (7 years, 5 months ago)

If you have to wipe with ISO, so be it... but why keep the burner(kkkj Jan*hner inside the SAB??.
Toutally un nessesary and dangerous. Just keep it  next to it.


--------------------

Amateur Mycologists United
AMU Q&A


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Invisible36fuckin5
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Re: print to agar [Re: 404]
    #23562335 - 08/21/16 08:24 AM (7 years, 5 months ago)

Quote:

404 said:
i wipe the insides of my SAB every time with iso. I also use the torch lighter inside with no problem. I can't for safety reasons suggest or condone this, but in my experience i have never had a problem and i've been doing it this way for several years.




Alcohol is a lot more expensive than soap and water. You're paying extra to make a fire hazard. And it will eventually blow up on you.


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


Extras: Filter Print Post Top
Offlinebokaj
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Re: print to agar [Re: 36fuckin5]
    #23562656 - 08/21/16 10:53 AM (7 years, 5 months ago)

i use tissue from inside the mushroom, the you get a mono culture.


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OfflinePantheist
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Re: print to agar [Re: bokaj]
    #23562827 - 08/21/16 11:58 AM (7 years, 5 months ago)

Thanks a lot for the responses everyone, some great reading - particularly in that thread with the naked guy who blew himself up and almost burnt the house down.

Quote:

mushpunx said:
There is no reason to wipe the insides with ISO.. its un nessacary and dangerous. If your SAB needs cleaning wash with dish soap and water.
You can't/dont need to anyways sterilize an SAB.

To prep a still air box you use a spray bottle of water with a little dish soap in it. You spray the walls lightly and the air a little and let it settle for a while. The soapy water will help trap airborne contams/settle the air quicker.

You do however, want to wipe all jars/plates down with ISO before placing in your SAB, and wash your gloves with ISO before and frequently during work.


To inoculate with a loop you would flame sterilize the loop untill red hot, cool it in the recieving blank agar plate, rub a tiny section of the print, and then swipe across the plate in a zig zag pattern.
When you cool the loop it gets some agar on it which helps to pick up spores off the platen




This info is a big help. One more question - if doing print to agar with an inoculation loop, can you just use one dab on the print to inoculate several petri dishes? If so, how many's a good number?


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InvisibleBunya
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Re: print to agar [Re: Pantheist]
    #23562882 - 08/21/16 12:20 PM (7 years, 5 months ago)

I sterilize between every dish.
If there's a contam on your first you will spread it to all the other ones if you dont sterilize between.


--------------------
Bunya cones kill.


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Invisible36fuckin5
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Registered: 08/11/03
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Re: print to agar [Re: bokaj]
    #23562945 - 08/21/16 12:47 PM (7 years, 5 months ago)

Quote:

bokaj said:
i use tissue from inside the mushroom, the you get a mono culture.




No, that can and almost always will sector. You'd still have to isolate if you want a pure culture.


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


Extras: Filter Print Post Top
Invisible404
error
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Registered: 08/20/10
Posts: 14,539
Re: print to agar [Re: 36fuckin5]
    #23562954 - 08/21/16 12:50 PM (7 years, 5 months ago)

Quote:

36fuckin5 said:
Quote:

404 said:
i wipe the insides of my SAB every time with iso. I also use the torch lighter inside with no problem. I can't for safety reasons suggest or condone this, but in my experience i have never had a problem and i've been doing it this way for several years.




Alcohol is a lot more expensive than soap and water. You're paying extra to make a fire hazard. And it will eventually blow up on you.





What? It's a dollar something at the store. Also, if it were going to blow up on me or catch fire it would have done so long ago i think, not that alcohol isn't flammable or anything... I've even had it pool on the bottom and used the flame inside and nothing happened. Typically tho, and i left this out, i wait a while to evap.


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Invisible36fuckin5
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Registered: 08/11/03
Posts: 12,079
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Re: print to agar [Re: 404]
    #23562959 - 08/21/16 12:53 PM (7 years, 5 months ago)

Water is a few cents per gallon and soap is dirt cheap. If you're letting your alcohol evap it's not there to do its' job. When you do a big operation these costs become obvious.


--------------------
Redd Foxx said:
If you're offended I don't give a shit and don't come see me no more.

Pat The Bunny said:
A punk rock song won't ever change the world, but I can tell you about a couple that changed me.

bodhisatta said:
i recommend common sense and figuring it out.

These are the TEKs I use. They're all as cheap and easy as possible, just like your mom.


Extras: Filter Print Post Top
Invisible404
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Registered: 08/20/10
Posts: 14,539
Re: print to agar [Re: 36fuckin5]
    #23562965 - 08/21/16 12:55 PM (7 years, 5 months ago)

No doubt


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OfflinePantheist
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Registered: 06/05/16
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Re: print to agar [Re: 36fuckin5]
    #23563051 - 08/21/16 01:32 PM (7 years, 5 months ago)

Quote:

36fuckin5 said:
Water is a few cents per gallon and soap is dirt cheap. If you're letting your alcohol evap it's not there to do its' job. When you do a big operation these costs become obvious.




Is it best to use distilled soapy water, or will soapy tap water suffice?


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Offlinemushpunx
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Re: print to agar [Re: Pantheist]
    #23563117 - 08/21/16 02:08 PM (7 years, 5 months ago)

Soapy tap water is fine.


Search the forum, you will find at least a couple people who got burned pretty bad when ISO vapors on thier SAB ignited.


--------------------

Amateur Mycologists United
AMU Q&A


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