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OfflinePsilocyBen17
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Rinsing Pins before dropping on Agar: Clarification Needed
    #20259804 - 07/11/14 08:38 PM (10 years, 6 months ago)

Hey guys,

Whipped up 10 plates of agar yesterday and its ready to go now. I have both invitro fruits and pins ready to use. I've been told its wise to rinse your tissues before putting them on agar. Can I just rinse them in a cup in my SAB? Does the water NEED to be sterilized or is tap water fine to use?


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OfflinecronicrMFacebook
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: PsilocyBen17]
    #20259813 - 07/11/14 08:40 PM (10 years, 6 months ago)

invitro fruits or pins don't bother, just drop them on the plate


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It doesn't matter what i think of you...all that matters is clean spawn
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OfflinePsilocyBen17
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: cronicr]
    #20259822 - 07/11/14 08:41 PM (10 years, 6 months ago)

FUCK YA DUDE!!! Congrats on the TAG!!!!  :hug:


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OfflinePussyFart
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: PsilocyBen17]
    #20259824 - 07/11/14 08:42 PM (10 years, 6 months ago)

Pins can be placed directly on the agar. The mycellium is rapidly growing and will outrun most contams.

That being said, the pin might roll around and contam the whole dam dish....ive never had luck with pins to agar for this very reason.....

Instead I usually tear them open and take a tissue sample from the inside of a fruit/pin with a sterile scalpel.

Don't rinse the pin in anything....it would just add another vector for contams.

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Invisible36fuckin5
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: cronicr] * 1
    #20259830 - 07/11/14 08:42 PM (10 years, 6 months ago)

Who the hell told you that?

Don't rinse any tissue samples. Drop a whole pin on agar, or rip the flesh of a mature fruit and take some from the middle.


--------------------
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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OfflinePussyFart
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: 36fuckin5]
    #20259836 - 07/11/14 08:43 PM (10 years, 6 months ago)

And the TC attack begins lol......











:excellent:

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OfflinePsilocyBen17
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: 36fuckin5]
    #20259842 - 07/11/14 08:44 PM (10 years, 6 months ago)

Thx guys, wish me luck.


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OfflinecronicrMFacebook
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: 36fuckin5]
    #20259844 - 07/11/14 08:44 PM (10 years, 6 months ago)

YEAH SAVE RINSING FOR DIRTY ASSS WILD FRUITS, THERE YOU CAN USE STERILIZED WATER TO WATER THE CONTAM RATE DOWN BUT FOR INDOOR PURPOSES IT'S A BACKWARD STEP..fucking caps lock lol


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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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Offlinesrphatness
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: 36fuckin5]
    #20259859 - 07/11/14 08:47 PM (10 years, 6 months ago)

So I guess that raises the question. Which is better (higher success rate). Cloning from pins or cloning from interior tissue of mature fruit?

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OfflinePussyFart
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: srphatness]
    #20259864 - 07/11/14 08:47 PM (10 years, 6 months ago)

It all depends on the genetics at hand....which is mostly always a mystery.

I have had success doing both.

Edited by PussyFart (07/11/14 08:48 PM)

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Invisible36fuckin5
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: srphatness]
    #20259866 - 07/11/14 08:48 PM (10 years, 6 months ago)

It's not about success rate, both will be 100% when done right. Tissue from younger samples will always hit senescence later than tissue from an older sample.


--------------------
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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OfflinecronicrMFacebook
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: 36fuckin5]
    #20259876 - 07/11/14 08:50 PM (10 years, 6 months ago)

yeah but with a mature fruit you knida know what your getting, pins can sorta be a guessing game but at the very least you know ya got a clue as to what you can expect(clusters,fast pinner that sorta thing)


--------------------
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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OfflinePsilocyBen17
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: cronicr]
    #20259886 - 07/11/14 08:51 PM (10 years, 6 months ago)

Im cloning a GIANT fruit tonight, hope it goes well.


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OfflinecronicrMFacebook
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: PsilocyBen17]
    #20259894 - 07/11/14 08:52 PM (10 years, 6 months ago)

:firstladyofapproval: what flush you cloning from?


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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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Invisible36fuckin5
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: cronicr]
    #20259909 - 07/11/14 08:54 PM (10 years, 6 months ago)

Quote:

cronicr said:
yeah but with a mature fruit you knida know what your getting, pins can sorta be a guessing game but at the very least you know ya got a clue as to what you can expect(clusters,fast pinner that sorta thing)




True. And IDK what the difference really is when it comes to hitting senescence. I've cloned pins and mature fruits, and I've gotten 2+ years out of both. If there's a real difference, it'd probably be weeks or a month or 2 tops. But there is a difference, and since I always shoot for optimal, I advise cloning younger specimens.

It's still better than isolating from spores and testing each one. You waste a lot of substrate on testing crap genetics that way.


--------------------
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
OfflinePsilocyBen17
Pin Pornographer


Registered: 10/20/13
Posts: 3,751
Last seen: 2 years, 29 days
Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: cronicr]
    #20259919 - 07/11/14 08:56 PM (10 years, 6 months ago)

second, but this tub didnt really have distinct flushes because most of the fruits grew invitro. I also have several invitro fruits in a BRF jar im going to use too....

Plus some spores. Im just going to have a combo party and PC another 10 plates tommorrow to do transfers....

BTW I made 20 plates and PC'd 10 last night to use in a few mins....

How long will the other 10 stay good for? Theyve been sitting in a cool spot and will get PC'd tmrw. any special procedures i should take if waiting loger then 24 hrs to use them.


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Invisible36fuckin5
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: PsilocyBen17]
    #20259949 - 07/11/14 09:01 PM (10 years, 6 months ago)

You mean no-pour agar jars? I would PC them right now. Agar isn't a place where you can cut any corners whatsoever.


--------------------
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
OfflinePsilocyBen17
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Registered: 10/20/13
Posts: 3,751
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: 36fuckin5]
    #20259957 - 07/11/14 09:02 PM (10 years, 6 months ago)

ok thx 36. So ALWAYS PC them as soon as you pour them?


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Invisible36fuckin5
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: PsilocyBen17]
    #20259971 - 07/11/14 09:05 PM (10 years, 6 months ago)

If you're gonna do no-pour, yes. You MAY get away with letting them sit out overnight, but I wouldn't fuck around here.

My suggestion is always to sterilize your jars separately and pour them, though. Not only can you avoid most of your condensation this way, but you get experience pouring agar without having to throw away petris that you fucked up.


--------------------
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
OfflinecronicrMFacebook
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: 36fuckin5]
    #20259977 - 07/11/14 09:06 PM (10 years, 6 months ago)

yes do it asap! i've waited less then 24 hours and got cbweb in half of them lol


--------------------
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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InvisibleMykes logos
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: PussyFart]
    #20259983 - 07/11/14 09:07 PM (10 years, 6 months ago)

Quote:

PussyFart said:
Pins can be placed directly on the agar. The mycellium is rapidly growing and will outrun most contams.

That being said, the pin might roll around and contam the whole dam dish....ive never had luck with pins to agar for this very reason.....

Instead I usually tear them open and take a tissue sample from the inside of a fruit/pin with a sterile scalpel.

Don't rinse the pin in anything....it would just add another vector for contams.




Alternatively, you could also use a small ruler to measure pins (or eye them out) and only cut pins that will touch the lid of the petri and wedge themselves between the agar and the lid.

Using very consistently poured triplates for this works well and saves dishes and parafilm...

cheers.

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Invisible36fuckin5
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: Mykes logos]
    #20259995 - 07/11/14 09:09 PM (10 years, 6 months ago)

Quote:

Mykes logos said:
Quote:

PussyFart said:
Pins can be placed directly on the agar. The mycellium is rapidly growing and will outrun most contams.

That being said, the pin might roll around and contam the whole dam dish....ive never had luck with pins to agar for this very reason.....

Instead I usually tear them open and take a tissue sample from the inside of a fruit/pin with a sterile scalpel.

Don't rinse the pin in anything....it would just add another vector for contams.




Alternatively, you could also use a small ruler to measure pins (or eye them out) and only cut pins that will touch the lid of the petri and wedge themselves between the agar and the lid.

Using very consistently poured triplates for this works well and saves dishes and parafilm...

cheers.




That sounds like it'd be a cunt. I always just cool my scalpel in the middle of my agar so it make a divot to put my sample in. Be careful with your dish (which you should anyway, it's lab equipment) and you shouldn't have issues.


--------------------
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
OfflinePsilocyBen17
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Registered: 10/20/13
Posts: 3,751
Last seen: 2 years, 29 days
Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: 36fuckin5]
    #20260019 - 07/11/14 09:14 PM (10 years, 6 months ago)

thx for the tips guys. one more question.

Im taking an inner sample from a big fruit. Do I tear it open inside my SAB or outside my SAB? If I toss the whole fruit into my SAB to tear it open wont it bring all the contams in with it?


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OfflinePussyFart
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: PsilocyBen17]
    #20260048 - 07/11/14 09:20 PM (10 years, 6 months ago)

Inside the SAB....you don't want the tissue touching open air for any reason.

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Invisible36fuckin5
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: PussyFart]
    #20262075 - 07/12/14 09:24 AM (10 years, 6 months ago)

Your SAB is already full of contams. You don't need a sterile environment, you need still air.


--------------------
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
InvisibleMr. Alien
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: cronicr]
    #20262098 - 07/12/14 09:30 AM (10 years, 6 months ago)

I had no luck with pins, mold and bacteria grow first, myc never appears.


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InvisibleMykes logos
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Re: Rinsing Pins before dropping on Agar: Clarification Needed [Re: Mr. Alien]
    #20262499 - 07/12/14 11:33 AM (10 years, 6 months ago)

Quote:

Mr. Alien said:
I had no luck with pins, mold and bacteria grow first, myc never appears.






I see myc... is this day 2 or 3?

EDIT: has anyone ever used H2O2 in spray form? I've tried it with clones, even pins, and it worked magnificently

Edited by Mykes logos (07/12/14 11:34 AM)

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