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#381 Mycol

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Posted 21 June 2020 - 11:39 AM

Hey everyone...this thread to get here is super long...sees to have drifted from spore vendors to all manner of useful info...
But I have a question....or three...
I just got my order from Sporeworks as they rated highly here and my original spores vendor in CO went out of business.
I got 2 10cc’s spore syringes...1-b+ and 1 called Puerto Rican* both are cube strains.
Looking at the contents, I notice a large mass of dark spore material and lots of floater pieces. Shaking it just moves it around.


“How can I ensure the clumped spore material will all come out of the syringe EVENLY?”
Thanx folks...help from folks here, and some in particular, have gotten me this far...Into my 2nd *project.
Peace all
P~

My Peurto ricans are not heavy sporers, so that might be why you don’t see much in there but mine are from SW too and doing well.

Shake the crap out of the syringe while turned sideways for as long as it takes to break them up into clumps. There should be a bubble in the syringe that moves back and forth to agitate them .

If that doesn’t work I’ve seen people squirt the solution into a brand new ziplock bag and suck it back up . The inside of the bag should be sterile . Never tried it but I have read it .

Also between every injection point give it another shake .

Edited by Mycol, 21 June 2020 - 11:41 AM.

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#382 Paradoc

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Posted 27 June 2020 - 06:51 AM

ThanxMuch, mycol...I appreciate your input. I finally opted to use another -smaller, Sterile 5cc syringe & 18ga. Needle and after shaking the original syringe like mad, inserted my smaller-New 5cc syringe with NEW needle, INTO the larger 10cc spore syringes I ordered. I inoculated my jars with this , 5cc syringe-(still cloudy, but...)
I had to ensure sterility, and having experience as a medico, I knew the 16ga needle should e a large enough bore (hole) to get the job done-as it’s likely to pass some *particulate matter through it.
*I probably didn’t even need to do this, but am glad I did.
*Again; ALL sterile procedures must be followed if you’re gonna do this, to minimize bad things getting into your world...

All the jars inoculated with it being done this way, are going well. I’m noticing fairly even colonizing around the jars-So I think I may have done this o.k. All this exercise really did was *assure* me I would inoculate my brf jars Evenly...
Your mention of emptying the Large syringe into a bag isn’t unique.
My direct observation was that in order to minimize any chance of any outside contaminant entering the picture, Direct Removal with a sterile needle worked well for me as I ‘could’ ensure the sterility of my syringe, but really ‘could not’ ensure, sterility for just about anything else....but I tend to overdo the front end of all this because I’m only just learning how all this goes.

Peace~
Thanx.
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#383 MycoAnt81

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Posted 04 July 2020 - 07:39 PM

One key thing I noticed is that many spore syringes are heavily contaminated.
In my experience 95% of them have bacterial or mold contam.
If you use them directly on grain, the loss ratio is too high.
Whenever I get a new syringe, I always test it on agar first.
In most cases, I do not go to grain until I have a viable culture on agar.

Curious what experience other have with spore syringes?

I’ve had great success with syringes. If I’m meticulous with my grain prep, I have 85% colonization, sometimes if all goes well. I don’t have a flow hood and the SAB didn’t help. Sporeworks has great products, just never had any luck with Penis Envy, EVER!!! Anything Mexican, B+ and Hawaiian I’ve had awesome results after a good grain prep.


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#384 Saphroziac

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Posted 05 July 2020 - 03:18 AM

Weird. I order PE from Sporeworks sometimes, and I've never even gotten it to colonize millet or hard red winter wheat berries. My azure syringe from them was contaminated. Happens sometimes I guess, I will still order from them 100%.


Edited by Saphroziac, 05 July 2020 - 03:18 AM.


#385 FunG

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Posted 05 July 2020 - 11:03 AM

Sporeworks has never sent me a contaminated syringe. They're pretty good.

I make my own from prints and always test the the syringe by inoculating a single jar of grain and not shaking it. That way when germination happens you can monitor where the solution has made contact with the grain for any signs of contamination....exactly as you would do with agar. The only downfall is you cant begin isolating out clean myc by testing on a grain jar but 99% of the time its clean growth anyways so pffft one way or the other.
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#386 Paradoc

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Posted 06 July 2020 - 10:23 AM

Heya, FunG...I’ve gotta Question about which I’m hoping you, or anyone else reading this, may have input and advice.
I’m really new at all this But have been as meticulous & careful as possible around spores. My SW project is rolling right along.
My project now has allowed me to collect spore prints on tinfoil. I put a small glass on top of it & wrapped foil around it.
*i am using Pre Made brf-type jars and I’m 23 for 24 using pre made spore syringes. (My error caused The 1 Failure)
*HOW would I make a spore syringe from my prints and not use agar?
I put them all the into the freezer, inside a new ziplock bag.

I think I read somewhere about literally taking the small pieces of foil, put it into a clean plastic bag...add water and free up the spores by agitating the whole thing from the foil. Draw them up into the syringe. Live long & prosper

Again, I’ve no idea where I saw this, nor even if it would work. Any comments are greatly appreciated & needed.

To master this particular step would, for me, complete the Circle of Life....and boy O boy, would I love to do that.

Thanx all. Be well
Peace~

#387 TVCasualty

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Posted 09 July 2020 - 09:42 AM

I make prints on foil cut to be slightly larger than the caps, then after letting them sit (covered, and usually in front of my flow hood) overnight to allow excess moisture to evaporate I vacuum seal them in foodsaver bags. NOTE: I don't vacuum the bags when I seal them, I only seal them. I make custom-size bags that are just slightly larger than the foil, which is helpful when it's time to put the spores in a syringe. I store my dry prints in the fridge after double-bagging them in large ziplock bags so I can take the bags off in layers; the outer bag is discarded at the door of the lab and the inner is opened in front of my flow hood.

 

 

To make syringes, I PC the empty syringes (wrapped in foil) and a jar of water. When the water has cooled to room temp I unwrap a syringe, open the jar of sterilized water, fill the syringe, then re-flame the needle until it's red hot and use the hot needle to melt a hole in a corner of the foodsaver bag. With the tip of the needle still in the bag, I push all the water out of the syringe into the bag in a forceful jet aimed at the spores to blast them off the foil. They can also be rubbed off the foil with your finger (rub the outside of the bag!). I then draw the spore water back up, cap the needle, and go on to the next one. I usually use 60cc syringes, so I make one syringe per print (fewer steps, less work than filling multiple 10cc syringes). At one print per syringe, the water in my syringes ends up being totally black/opaque with spores, a luxury you can afford when you make your own.

 

Go through all of the motions in a full dress rehearsal (use blank "prints") before doing it for real the first time so you know what to expect without risking a print. One of the first things you'll notice is that you need three hands to pull this off, but you can get away with two and a half (I draw up the water by pulling the syringe back with my teeth while holding the bag in one hand and controlling the placement of the syringe with the other). I've been meaning to improve this procedure since I'd prefer to not involve my mouth in tissue culture work, lol.

 

I try to poke through the bag so the (blunt!) needle hits the foil which helps prevent it from melting through the other side (making two holes instead of one). If done carefully and quickly the syringes can be filled without having to sit in front of a flow hood. But if you have one then I'd definitely do everything in front of it anyway.

 

Hope that helps.


Edited by TVCasualty, 09 July 2020 - 09:44 AM.

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#388 Paradoc

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Posted 11 July 2020 - 01:00 PM

Wow. TV, Thanx. I’m working up the nerve to do this and am super grateful for your very detailed step by step instructions.
I’ll take your advice and do a *test run-
I don’t have a flowhood, but reading through this I think I now more clearly get the concept.
Doing this with vacuum sealed bags makes sense. I didn’t do that this time.
My prints were just stored on foil-in a lighter gauge plastic ziplock bag.

I appreciate your taking time to reply.
Peace
P~
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#389 Jrotten

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Posted 11 July 2020 - 02:24 PM

I know it's not the purpose of this thread, but is there an advantage to a spore syringe versus a liquid culture made from agar and a spore print?  Having recently taken to agar it just seems easier to keep and LC clean in the making as opposed to a spore syringe.



#390 TVCasualty

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Posted 11 July 2020 - 02:30 PM

I know it's not the purpose of this thread, but is there an advantage to a spore syringe versus a liquid culture made from agar and a spore print?  Having recently taken to agar it just seems easier to keep and LC clean in the making as opposed to a spore syringe.

 

I don't know, I guess it would depend on what you're planning to do after that initial step.


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#391 bwahaha

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Posted 28 July 2020 - 09:51 AM

anyone had experience microtech spore supplier

I feel i made a bad choice. Would like to know if re-inoculating the jars would be fine.its been a week and a day with no seen growth on rye. other vendor shows progress already.






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