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NumeroEno
I come from the land of lizards



Registered: 07/24/14
Posts: 9,652
Loc: Gamehendge
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First time ever growing from a print. Wanna make sure I'm about to do it right.
#20617298 - 09/25/14 01:52 PM (10 years, 3 months ago) |
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So I received a purple mystic print yesterday, which is a variety I've been really curious about. I have some sterile pasty plates ready to go, but I'm wondering what is the best way to get the spores on agar. My plan is to PC a syringe full of water, a shot glass, and a ceramic plate (to have as a sterile work surface) and put them in the SAB while still hot.
Once the air settles, I'm gonna sanitize the plate and then rinse the spores off into the shot glass with the sterile water from the syringe. Then I plan to suck everything back into the syringe, never having my hands directly over the print or the agar plates. Once the syringe is loaded with spores I'm gonna unwrap and inoculate the agar plates.
My questions are:
1. Should I flame the needle at any time since it's going into the SAB sterile?
2. Is the plate necessary? I was worried about the stability of the shot glass on the towels.
3. Is this spore syringe gonna be good for a long time, considering only a few drops of solution are going onto the agar plates?
4. Purple mystic is a pretty recently discovered variety. What will the best way to tame it? Should I start with just one plate and repeatedly transfer the most aggressive mycelium before inoculating grain with it?
I'm about to go to work and plan on doing this when I get home in 8 hours, so I'll take all the advice I can get in the mean time.
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Let it grow! Let it grow! Greatly yield!
What shall we say, shall we call it by a name
As well to count the angels dancing on a pin
Water bright as the sky from which it came
And the name is on the earth that takes it in
DOG FOOD AGAR
MY ELECTRIC INOCULATION LOOP
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Sagescruffy
CH



Registered: 10/30/09
Posts: 2,011
Loc: PNW
Last seen: 4 months, 3 days
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: NumeroEno]
#20617335 - 09/25/14 01:58 PM (10 years, 3 months ago) |
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Using an inoculation loop would be better than going the syringe route because it will be less likely for bacteria to be present, less vectors of contamination, and likely faster germination. If you put a drop of spore solution onto a plate of agar that drop of water will just sit there while the spores from a print would have all ready germinated.
-------------------- Love.  
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NumeroEno
I come from the land of lizards



Registered: 07/24/14
Posts: 9,652
Loc: Gamehendge
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: Sagescruffy]
#20617782 - 09/25/14 03:43 PM (10 years, 3 months ago) |
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Right on. I have some 24 gauge kanthal that would make a sweet inoculation loop. I could just stick it in a piece of wood I guess.
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Let it grow! Let it grow! Greatly yield!
What shall we say, shall we call it by a name
As well to count the angels dancing on a pin
Water bright as the sky from which it came
And the name is on the earth that takes it in
DOG FOOD AGAR
MY ELECTRIC INOCULATION LOOP
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ant707
not a puppet


Registered: 07/26/14
Posts: 579
Loc: northeast florida
Last seen: 7 years, 11 months
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: NumeroEno]
#20617800 - 09/25/14 03:46 PM (10 years, 3 months ago) |
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Quote:
EnoRoxxx said: Right on. I have some 24 gauge kanthal that would make a sweet inoculation loop. I could just stick it in a piece of wood I guess.
you'll get a lot more use out of your prints using a loop too! 
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: NumeroEno]
#20617803 - 09/25/14 03:46 PM (10 years, 3 months ago) |
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I've used paper clips, big safety pins, my scalpel, they all good. Just streak the spores in a line across the plate and then grab from the fastest growth you can see. If the print is wild you might need to be extra vigilant regarding contams.
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kmetric



Registered: 08/23/14
Posts: 140
Last seen: 8 years, 10 months
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: Sagescruffy]
#20617815 - 09/25/14 03:49 PM (10 years, 3 months ago) |
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Quote:
Sagescruffy said: Using an inoculation loop would be better than going the syringe route because it will be less likely for bacteria to be present, less vectors of contamination, and likely faster germination. If you put a drop of spore solution onto a plate of agar that drop of water will just sit there while the spores from a print would have all ready germinated.
I've heard this a lot..I just started working with agar and for me, spores straight to agar showed no growth (over 14 days..) while a single drop of spore solution showed growth on almost all plates. I've done a bunch of pastyplates too.
Weird... I was thinking my print is a bit old so rehydrating them first helped out. But they're only around a year old. *shrug*
Edited by kmetric (09/25/14 03:52 PM)
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: kmetric]
#20617871 - 09/25/14 03:58 PM (10 years, 3 months ago) |
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Sometimes if the spores are old, rehydrating them (ie in a syringe) will perk em up. You can also melt a puddle in the plate and then swipe the spores in that.
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NumeroEno
I come from the land of lizards



Registered: 07/24/14
Posts: 9,652
Loc: Gamehendge
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: Pastywhyte]
#20617941 - 09/25/14 04:08 PM (10 years, 3 months ago) |
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The spores aren't wild but the spores that started the previous generation were, apparently. It's a nice clean print AFAIK.
--------------------
Let it grow! Let it grow! Greatly yield!
What shall we say, shall we call it by a name
As well to count the angels dancing on a pin
Water bright as the sky from which it came
And the name is on the earth that takes it in
DOG FOOD AGAR
MY ELECTRIC INOCULATION LOOP
Edited by NumeroEno (09/25/14 04:09 PM)
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: NumeroEno]
#20618071 - 09/25/14 04:34 PM (10 years, 3 months ago) |
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It will be easy then just put em on the plate
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NumeroEno
I come from the land of lizards



Registered: 07/24/14
Posts: 9,652
Loc: Gamehendge
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: Pastywhyte]
#20618344 - 09/25/14 05:23 PM (10 years, 3 months ago) |
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Shweet.
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Let it grow! Let it grow! Greatly yield!
What shall we say, shall we call it by a name
As well to count the angels dancing on a pin
Water bright as the sky from which it came
And the name is on the earth that takes it in
DOG FOOD AGAR
MY ELECTRIC INOCULATION LOOP
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MrGiraffe

Registered: 04/04/14
Posts: 3,149
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: Pastywhyte]
#20618476 - 09/25/14 05:46 PM (10 years, 3 months ago) |
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Quote:
Pastywhyte said: Sometimes if the spores are old, rehydrating them (ie in a syringe) will perk em up. You can also melt a puddle in the plate and then swipe the spores in that.
 I like making a little divot in the center and then swiping spores into that divot. I always have better germination rates that way.
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NumeroEno
I come from the land of lizards



Registered: 07/24/14
Posts: 9,652
Loc: Gamehendge
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: MrGiraffe]
#20618646 - 09/25/14 06:20 PM (10 years, 3 months ago) |
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Cool. Just a few more hours until I'm off work and it's time to play with the print.
Thanks for the tips. Seems like you all saved me a lot of trouble. I guess I don't really ever need to make a syringe for any reason
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Let it grow! Let it grow! Greatly yield!
What shall we say, shall we call it by a name
As well to count the angels dancing on a pin
Water bright as the sky from which it came
And the name is on the earth that takes it in
DOG FOOD AGAR
MY ELECTRIC INOCULATION LOOP
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MrGiraffe

Registered: 04/04/14
Posts: 3,149
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: NumeroEno]
#20618652 - 09/25/14 06:21 PM (10 years, 3 months ago) |
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Quote:
EnoRoxxx said: Cool. Just a few more hours until I'm off work and it's time to play with the print.
Thanks for the tips. Seems like you all saved me a lot of trouble. I guess I don't really ever need to make a syringe for any reason 
Not if you've got agar and plates.
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: NumeroEno]
#20618654 - 09/25/14 06:21 PM (10 years, 3 months ago) |
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Quote:
EnoRoxxx said: Cool. Just a few more hours until I'm off work and it's time to play with the print.
Thanks for the tips. Seems like you all saved me a lot of trouble. I guess I don't really ever need to make a syringe for any reason 
I have never made a spore syringe. Never needed to.
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NumeroEno
I come from the land of lizards



Registered: 07/24/14
Posts: 9,652
Loc: Gamehendge
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: Pastywhyte]
#20618944 - 09/25/14 07:20 PM (10 years, 3 months ago) |
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Wonderful. It seemed like a PITA. I have a tendency to over-complicate things, especially when I drink a lot of caffeine. I was preparing myself for an hour long syringe mission, but instead this will be quick and easy. Now if I could only stop worrying about how I might have compromised my other project...
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Let it grow! Let it grow! Greatly yield!
What shall we say, shall we call it by a name
As well to count the angels dancing on a pin
Water bright as the sky from which it came
And the name is on the earth that takes it in
DOG FOOD AGAR
MY ELECTRIC INOCULATION LOOP
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NumeroEno
I come from the land of lizards



Registered: 07/24/14
Posts: 9,652
Loc: Gamehendge
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: NumeroEno]
#20619889 - 09/25/14 10:33 PM (10 years, 3 months ago) |
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Got home from work, burned a bowl, and built this guy. Looks like all the ones I've seen online except with a wood handle, which I sealed with loctite plastic adhesive. Good shit.
Any tips on technique, just scrape it off the print into the agar? Suffice it to say this thing gets heavily flame sterilized?
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Let it grow! Let it grow! Greatly yield!
What shall we say, shall we call it by a name
As well to count the angels dancing on a pin
Water bright as the sky from which it came
And the name is on the earth that takes it in
DOG FOOD AGAR
MY ELECTRIC INOCULATION LOOP
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MrGiraffe

Registered: 04/04/14
Posts: 3,149
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: NumeroEno]
#20620258 - 09/26/14 05:44 AM (10 years, 3 months ago) |
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Flame. Scrape. Swipe
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Pastywhyte
Say hello to my little friend



Registered: 09/15/12
Posts: 37,972
Loc: Canada
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: MrGiraffe]
#20620272 - 09/26/14 05:58 AM (10 years, 3 months ago) |
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You don't need a lot of spores, the tiniest visible spec is thousands.
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NumeroEno
I come from the land of lizards



Registered: 07/24/14
Posts: 9,652
Loc: Gamehendge
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: Pastywhyte]
#20620850 - 09/26/14 09:58 AM (10 years, 3 months ago) |
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I did it last night and I think it worked. I ended up getting a lot of spores on there, like the size of a pin head. Next time I wont apply any pressure on the loop while I'm scraping the print. I'm guessing what I did last night will work fine though. I also picked up some new ziploc containers for my other project, and they happen to be the perfect size for my scalpel handle and loop. I'll try to get some pics up once I see growth.
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Let it grow! Let it grow! Greatly yield!
What shall we say, shall we call it by a name
As well to count the angels dancing on a pin
Water bright as the sky from which it came
And the name is on the earth that takes it in
DOG FOOD AGAR
MY ELECTRIC INOCULATION LOOP
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kmetric



Registered: 08/23/14
Posts: 140
Last seen: 8 years, 10 months
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Re: First time ever growing from a print. Wanna make sure I'm about to do it right. [Re: NumeroEno]
#20620894 - 09/26/14 10:13 AM (10 years, 3 months ago) |
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Nice OP 
Speaking of applying pressure..do you guys think maybe none of my scraped plates germinated because I applied too much pressure and ripped up all the spores? That was a thought I had. That or I put way too many spores at once.
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