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Invisiblecattledog
Dog

Registered: 11/26/19
Posts: 32
Loc: Canada Flag
Crazy #s of contams in cloned dish. Is this typical?
    #26509923 - 02/29/20 03:46 PM (3 years, 10 months ago)

*

*

*55mm dishes

I cloned two parts of a mushroom into two different dishes. The widespread location of the contams leads me to believe I must have put my hand over the dish. Would that do it?

-An inexperienced agarer


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Teks 4 Success


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OfflineMYCELIUMINDED
Fungal phenomenon
I'm a teapot

Registered: 02/25/20
Posts: 24
Loc: Canada
Last seen: 3 years, 3 months
Re: Crazy #s of contams in cloned dish. Is this typical? [Re: cattledog]
    #26509926 - 02/29/20 03:47 PM (3 years, 10 months ago)

I haven’t done anything yet


But from
My reading of everything yes it would


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Starting from square 1
More like Petri dish 1
Lololololololo

Mushroom mayhem upon us


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OfflineA.k.aM
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Re: Crazy #s of contams in cloned dish. Is this typical? [Re: cattledog]
    #26509931 - 02/29/20 03:50 PM (3 years, 10 months ago)

Yeah you don’t want anything over the dish.

With cloning you have to be careful to peel the stem back without touching the insides and then cut a piece out without cutting into the outside of the stem. Only the tissue inside is clean any part that was  exposed to the air while growing will usually bring all kinds of contams.

Is that pin you put on the plate from another dish?

It takes a while but eventually you’ll automatically stop moving over your plates. Lol I’ve caught myself moving my hands and arms around pots instead of over while cooking and stuff like that. It just gets incorporated into your brain.


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LAGM2020


Edited by A.k.a (02/29/20 03:52 PM)


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OfflineIgnorantape
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Registered: 01/30/20
Posts: 149
Loc: The Indigo Plateau
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Re: Crazy #s of contams in cloned dish. Is this typical? [Re: A.k.a]
    #26509958 - 02/29/20 04:09 PM (3 years, 10 months ago)

Not necessarily your hands. That horizontal smear in the second pic looks like you swiped a dirty loop over the agar. It suggests that your sample was contaminated.

Can you describe the procedure that you did when you took the sample and laid it on the plate?


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Invisiblecattledog
Dog

Registered: 11/26/19
Posts: 32
Loc: Canada Flag
Re: Crazy #s of contams in cloned dish. Is this typical? [Re: A.k.a]
    #26511826 - 03/01/20 06:35 PM (3 years, 10 months ago)

Quote:

you have to be careful to peel the stem back without touching the insides and then cut a piece out without cutting into the outside of the stem.




There s a great chance this is where I screwed up. In trying to cut chunks out the stipe of one, I ended up mutilating it pretty badly and may have brushed the outside.

The pin I took is straight from a cake. I have no idea why I thought I could clone a pin grown in nonsterile air a week ago.

I aspire to develop your muscle memory with agar. Thank you.


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Teks 4 Success


Edited by cattledog (03/01/20 06:35 PM)


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Invisiblecattledog
Dog

Registered: 11/26/19
Posts: 32
Loc: Canada Flag
Re: Crazy #s of contams in cloned dish. Is this typical? [Re: Ignorantape]
    #26511832 - 03/01/20 06:40 PM (3 years, 10 months ago)

Good thinking.

This was my procedure:
1. Take my best mushroom into the SAB and tear it open (with my hands)
2. Hold it with one hand in the SAB and use a lighter to flame an Xacto outside the SAB until the tip turns red
3. Cool the Xacto in the dish
4. Cut out a piece from the stipe
5. Place it on the dish

I noticed you mention loop instead of a knife for getting a tissue sample. Is a loop the preferred (or necessary) tool?


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OfflineIfkilbadluck
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Registered: 03/22/18
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Re: Crazy #s of contams in cloned dish. Is this typical? [Re: cattledog]
    #26511936 - 03/01/20 08:12 PM (3 years, 10 months ago)

Quote:

cattledog said:
Good thinking.

This was my procedure:
1. Take my best mushroom into the SAB and tear it open (with my hands)
2. Hold it with one hand in the SAB and use a lighter to flame an Xacto outside the SAB until the tip turns red
3. Cool the Xacto in the dish
4. Cut out a piece from the stipe
5. Place it on the dish

I noticed you mention loop instead of a knife for getting a tissue sample. Is a loop the preferred (or necessary) tool?



1) all good
2)still all good but I learned a easier way when you are new is to have another Petri in the sab, and set the knife down on the Petri with the blade hanging off of the dish. Never let your hand go over the blade while cooling.
3) follow above and let it cool naturally. Also invest in a scalpel they are cheap and far superior. I bought one with 10 blades over a year ago when I was new for 10$ still haven’t had to buy more blades or a new Handel.
4)I go for the smallest amount of inner tissue as possible. Little speck does the trick.
5)make sure your hand never gets even close to going over your plate.
If the pin was from a tub not a agar dish that would explain the contamination issues. I would suggest wit future clones to flame the blade and let it cool naturally. Don’t cool in agar as it is another contamination vector and isn’t the easiest thing when you are new to grasp. I ran into issues doing that when I was new.


Edited by Ifkilbadluck (03/01/20 08:24 PM)


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OfflinecronicrFacebook
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Re: Crazy #s of contams in cloned dish. Is this typical? [Re: Ifkilbadluck]
    #26511981 - 03/01/20 08:54 PM (3 years, 10 months ago)

looks more like your scalpel then your hand, if you struggle to get your tissue off you may be knocking contams off your scalpel in the process, new sharp blades are my go to for cloning.
cloning puns off cakes is more than fine, contams are expected with clones the goal is to get the mycelium growing faster than the contams and transfer asap.


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It doesn't matter what i think of you...all that matters is clean spawn

I'm tired do me a favor


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Invisiblecattledog
Dog

Registered: 11/26/19
Posts: 32
Loc: Canada Flag
Re: Crazy #s of contams in cloned dish. Is this typical? [Re: Ifkilbadluck]
    #26513244 - 03/02/20 05:04 PM (3 years, 10 months ago)

Thanks for the advice. I will try your cooling on spare dish idea and will buy a scalpel. I did not know they were more effective than an Xacto.


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OfflineIgnorantape
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Registered: 01/30/20
Posts: 149
Loc: The Indigo Plateau
Last seen: 3 years, 1 month
Re: Crazy #s of contams in cloned dish. Is this typical? [Re: cattledog]
    #26513331 - 03/02/20 05:58 PM (3 years, 10 months ago)

I didn't read properly. I thought you were laying spores rather than taking clones. Scalpel is the right thing to use for that.


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