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OfflineiEnter
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How is Everyone Doing Spore Swabs (APE) to Agar These Days?
    #26329287 - 11/18/19 05:04 PM (4 years, 2 months ago)

I've been using an inoculation loop to put all my spores to agar, off of prints and LC.

I just got 2 swabs of APE in, and I want to make sure I'm up to date before I get into this new territory.

Is it possible to just pull some spores off the swab with an inoculation loop?



I found this TEK which says:

Quote:

The TEK:
1. Position all your items in front of the flowhood or in an SAB.
2. If you are working with a stubborn print, first make a swab from the print. Try to collect the spores to one side.
3. Cut a small square or circle from the center of the agar. It is best to be just slightly larger than the tip of the swab and to not cut all the way through the agar (make a well in the center of the dish).
4. Carefully position the swab tip in the center of the well you just made, then cut it with the scissors such that it falls into the center of the well (preferably spores downward- you can use a sterile scalpel or sterile forceps to position the swab tip as you'd like.
5. Carefully soak the swab tip with sterile water. Having some excess water in the bottom of the well is fine, but you want to eliminate the ability for water to slosh all around the plate.
6. Wrap and let sit agar side down until germination.
7. Proceed with transfers :smile:




If this is how people are doing it these days, is cutting a "well" into the center and filling with water necessary? It seems to me that if I just cut the swab tip and left it on the agar, it would be less chance of contam, but take longer (which is cool with me). 


Other than that thread, most of the stuff I found through the search deal with using swabs to take spores from prints and LC to agar


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Invisiblenatedawgnow
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: iEnter]
    #26329612 - 11/18/19 06:40 PM (4 years, 2 months ago)

Dont do that tek. Swipe the swab directly to agar or if nothing grows use your
sterile scalpel to grab a few spore laden fibers and press them into your agar.
Prep your agar soft but not too soft


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OfflineCamera93
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: natedawgnow]
    #26329850 - 11/18/19 08:29 PM (4 years, 2 months ago)


:whathesaid: would be my 1st route.
I finally had luck just plunging the swab into a no pour.

This was the resulting transfer after germination


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OfflineiEnter
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: natedawgnow]
    #26330425 - 11/19/19 08:00 AM (4 years, 2 months ago)

Quote:

natedawgnow said:
Dont do that tek. Swipe the swab directly to agar or if nothing grows use your
sterile scalpel to grab a few spore laden fibers and press them into your agar.
Prep your agar soft but not too soft




For sure, Thank you! I'm going to swipe a couple plates with the first one. I'll up date on how that goes.

If it doesn't work I'll try some other ways.

I'll keep this thread updated in case anyone else is in my shoes.


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OfflineFailboat
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: iEnter]
    #26330471 - 11/19/19 08:21 AM (4 years, 2 months ago)

Be sure to use one swab on multiple plates so as to get the best bang for your buck and spread the spores out for growth selection. Save the second swab inccase the first fails.


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OfflineiEnter
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: Failboat]
    #26333573 - 11/20/19 01:22 PM (4 years, 2 months ago)

Quote:

Quirkmeister92 said:
Be sure to use one swab on multiple plates so as to get the best bang for your buck and spread the spores out for growth selection. Save the second swab inccase the first fails.




That's exactly what I did.

I swabbed 3 plates, then I just decided to break off the swab and stick it 1/2 way into the middle of the 4th plate.

I'll update on how they are going when I see growth.


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OfflineKizzle
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: iEnter]
    #26333608 - 11/20/19 01:41 PM (4 years, 2 months ago)

Quote:

Is it possible to just pull some spores off the swab with an inoculation loop?



Maybe. I think next time I'm actually going try forgetting the swabs altogether and collecting the spores with loops.


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OfflineOne of Us
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: iEnter]
    #26333630 - 11/20/19 01:54 PM (4 years, 2 months ago)

I was wondering how this was done as well. I have one swab each of two really cool strains I would love to grow out. I'm kinda worried about screwing it up and losing the genetics.

They are fresh spores though, so it's probably no big deal. I am probably hesitating for no real reason


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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: One of Us] * 1
    #26333909 - 11/20/19 03:59 PM (4 years, 2 months ago)

I go from syringe with 3 drops on sterile q-tip to streak onto agar with consistent results.  APE in sig.


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OfflineMycoactive
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: StoneJackBaller]
    #26333954 - 11/20/19 04:25 PM (4 years, 2 months ago)

It depends on how many swabs you have. If it's just one and you want to make sure you're preserving the genetics, you can use an inoculation loop as you suggested or even cut small pieces off onto the agar with sterile scissors and preserve the rest of the swab. If you have multiple swabs and/or don't care about the risk of losing the genetics, the easiest way to do it, as far as I know, is to wipe the swab directly on the agar. The efficiency isn't great in that you won't come close to getting all the spores off of the swab, but you should get plenty for myc growth.


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OfflineiEnter
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: Mycoactive]
    #26336843 - 11/21/19 07:53 PM (4 years, 2 months ago)

Quote:

It depends on how many swabs you have. If it's just one and you want to make sure you're preserving the genetics, you can use an inoculation loop as you suggested or even cut small pieces off onto the agar with sterile scissors and preserve the rest of the swab. If you have multiple swabs and/or don't care about the risk of losing the genetics, the easiest way to do it, as far as I know, is to wipe the swab directly on the agar. The efficiency isn't great in that you won't come close to getting all the spores off of the swab, but you should get plenty for myc growth.




I'm down to 1, I'm going to try this with this last one:
Quote:

Personally I take some sterile scissors and cut a piece of the spored cotton away from the swab, leaving most of the spores on the swab. Then I take the piece I cut off and grab it with sterile tweezers and swipe it across the agar.

This avoids getting agar on your swab and having spores germinate on your swab



Found it Here

Cutting off but then doing a couple dishes with it using tweezers

Then I'm going to use a loop on what's left

Nothing showing on the first set of plates yet, but I'll update on how they all do.


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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: iEnter]
    #26336949 - 11/21/19 08:36 PM (4 years, 2 months ago)


Disregard anything asking you to add water to hydrate a swab. Just throw that out. It’s too complicated. I make my own. Sterile Swabs Now

I find swabs much more friendly than spore loops. You don’t have to flame anything either. Just swipe the plate all over with the end of the swab then chunk it. I use swabs to start all of my prints on plates now. I won’t go back either. Easy as cake.

I’ve also had my presterilized swabs contained in a mason jar for close to two years. I’ve started 20 plus plates this year without one contam. I used a sab and pasty plates. Wipe the top of the lid with alc. loosen the lid. Turn the pasty plate 90 degrees. Loosen lid  then swab the plate. I use the lid and plate as a shield so things from above don’t fall on the swab or agar. I use circular or zig zag motion as I swab. I also twirl the swab as I am swiping the spores in the plate. So it’s a working motion. But subtle.

What is so great about the way I wrap each swab with foil is that it is contained when you place it in the sab. Then you only unfold it right before you do the swab. I also fold my prints where, when they unfold, there is a foil flap that is lifted above the print that also acts as a shield from thing falling from above. So when I swipe that swab on the print the print is covered by the top foil. Sometimes I even use the technique of flipping the print upside down and taking a swipe upside down. I then have my plate close by with loosened lid that I swab. Steady motions. Not to aggressive.


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Edited by eatyualive (11/21/19 08:55 PM)


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OfflineiEnter
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: eatyualive]
    #26339225 - 11/22/19 10:00 PM (4 years, 2 months ago)

Awesome tek man! I'll be swabbing the APE once I get them going.

I'll give this a shot then


Update: First set of plates (rubbing swabs on agar and cutting off entire cotton and leaving it) are showing. I'll get some pics in a few days when I get back with them!


Edited by iEnter (11/22/19 10:05 PM)


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OfflineiEnter
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: iEnter]
    #26365136 - 12/06/19 01:54 PM (4 years, 2 months ago)

Both ways work.


Here are the 3 that I swabbed the spores onto:







I believe first one was a bit discolored in one spot (can't see it because of my god awful camera)

The second was perfect

Idk what happened to the third lol



Here is the one that I left the cotton tip on:

Top



Bottom


You can see there is something growing under the myc right where I left the swab



So I took a few cuts from all of them and now have 6 fresh APE plates.

Both ways work.


When I end up using that second swab I will try cutting pieces of cotton off and post up here.


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Offlinetrippleblack
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: iEnter]
    #26365145 - 12/06/19 02:00 PM (4 years, 2 months ago)

When i did ape agar work, i rubbed a swab with a metal inoculation loop..  I also pressed a swab on agar before, but didn't like the extra agar which got left on the swab.  least favorite, but worked well, was taking a pair of tweezerse and  pulling a few fibers off the swab..

either technique, there was no problems getting germination.


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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: trippleblack]
    #26365693 - 12/06/19 06:40 PM (4 years, 2 months ago)

I just germinated some ape on agar the other day. Never played with a swab before so I just stabbed the damn thing into the agar and left it lol. Got mycelium going good, so that's a tek too haha.


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OfflineReliableSporesV
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Re: How is Everyone Doing Spore Swabs (APE) to Agar These Days? [Re: iEnter] * 1
    #26365721 - 12/06/19 06:53 PM (4 years, 2 months ago)

No.  Do not cut a hole in the agar.  Well-hydrated spores scraped with a loop and then lightly scored onto the top of the agar is enough.  Less is more.  If the spores are viable this is enough.  It's easy to make this complicated real fast but it won't increase your chances of success. 

With agar you don't have to go overboard with precautions to prevent contamination unless you're looking for 100% clean dishes.  In agar, contaminants will present themselves quite clearly so the isolation of the good mycelium is easy.

There are ton of ways to overthink mycology.  Don't do it.  These guys have been around WAY longer than us and only need a sliver of opportunity to do their thing.  Just play the odds when inoculating anything, be it agar or grain.  Do more than you need and accept the failures as part of the process.  It's like trading stocks.  You let the winners run and cut the losers ASAP.


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