71 replies to this topic

[chill]

Hi all,

Bluehelix mentioned something called a vacutaner for storing liquid cultures.

That got me thinking that wouldn't it be possible to use steralized 4% sugar water to make spore syringes. That way the spores would germinate while they are in the syringe and would begin to immediately colonize the jar when injected. This would be helpful if you have a few minutes to make some syringes but plan on doing the jars later in the week for example.

So I tried this using maple syrup about a week ago and haven't noticed any growth in the syringe. I plan on innoculating wednesday and will see how it goes.

Has anyone else tried this and does anyone have any suggestions as to how to go about generating growth inside a syringe?

[motograter]

I was thinking about this today too. Why not just make up a jar for LC the normal way, then take a syringe that you only have a small amount left in, then suck up the LC? I'm going to try it this way tomorrow. (All out of Vanilla....)
Freak, keep us posted on your attempts!
I'll do the same

[nepenthes_ak]

I was just thinking about this the other day... hmm yes please let us know.

Cheers

[chill]

My motivation for doing the LC in the syringe is because I have bad comtam issues with LC jars.
The syringe should be airtight and thus safe against contams.
'Freak, you may want to incubate the syringe rather than letting it sit at room temp. I think you've mentioned that your place is cold and cool temps would stop a syringe from otherwise germinating.
Cool, thanks for your input guys. I think this could really speed up the process.
I'd like to get the total time from innoculation to cropping down to a month if possible.

[anticheffy]

Im thinking the air tight thing would work against you.......

[chill]

I just checked my spore stash and I have a couple of prints left over.
They are GT and very nice fruiters.
I have a proposition for someone. I will donate a GT print to the first person to successfully create an 'invitro' LC syringe. The spores and water must be mixed together in the syringe and kept there until mycelium start to grow. Pics will be required.
I'll mail out the print once the tek has been verified by another board member. I'll also send out a print to that person (kind of like an assist in hockey) :)
I'm kind of low on spores or I'd offer more but I think this is a fun way to get things rollin'.

[chill]

Yea, that is a possibility. I've tried introducing an air bubble into the syringe as one way of dealing with it.

Also, I've seen mycelium in a syringe before and hip has commented on this before so I am hopeful. I believe it is possible.

Im thinking the air tight thing would work against you.......

[anticheffy]

theres a pic of a myc syringe somewhere here that has what appears to be a sclorotia grown out in it, it was so old.....I cant find it though

[chill]

Hi 'freak,
I forgot about that tek with the colonized jar. Thanks for the reminder.
'Course you have to have a jar sitting around to use and this way you could just have a print (like I have) :)
I think I'll try some potatoe water this weekend. I had OK results with this one time.

[chill]

I boiled my water to kill any contams. I was wondering if boiling removes O2 so I did a little research.

I found this little nugget on a gov't web site:
"The solubility of oxygen and other gases depends on water temperature. The colder the water, the more gases it can hold. Boiling water removes all gases."

Maybe reverse osmosis water would be better to use?

Also, shaking the water appears to put O2 back into it. Possibly having a bubble in the syringe and then shaking it would do the trick.

[BuckarooBanzai]

Perhaps leave a good bit of air (1/4 of it) and toss in a BB or two? That way you can shake the syringe to keep oxygen in the solution and break up the developing mycelia.

I believe air tight will work as long as you leave a good bit of air inside the syringe. The LC will need some air.

I totally agree with using a 60mL syringe, too.

[chill]

Have been thinking about the heat and air thing.

I decided to get some 'clean' water (reverse osmosis) and make a syringe out of it without heating it.

I bought a one litre bottle, dumped out half of it, shook it up real good to put some air in it (just in case).

Then I added 25ml of maple syrup and shook that up real good. I then pushed a syringe needle into the bottle and sucked up liquid to use in making spore syringes.

One syringe is a 30cc sucker that I have left 10cc of air in for shaking purposes. The other is a regular 10cc.

I've got two syringes in total down now for incubating. Hopefully this will show some results.

I used my first attempt from last week to innoculate some GT jars today. We'll see how they perform compared to the regular syringes I also used as side-by-side.

[motograter]

I just started 1 syringe, I had a little under .5cc of GT water, made up a LC jar, and sucked up 8cc of that. I made sure to leave enough room to shake, a little. now its in my inc. chamber. We'll see in a week or 2 if its working out.

[chill]

Cool,

looks like we may have the first winner.

Pix and a mini tek write up would be sweet.

I'm going to refrain from checking on my syringes until at least 24 hours have passed.

[arezap]

Im thinking the air tight thing would work against you.......

Actually anticheffy, faof made some vLC's with 3 holes that were plugged with silicone caulk. faof pc'd the jars with toothpicks under the lids and allowed them to cool overnight to avoid cooling air in the jar creating a vaccuum. When opened the pc was opened the jars were sealed tightly. To load a syringe faof flames the needle and draws sterile air in the syringe, lets the needle cool, injects the sterile air into the jar, turns the jar upside down, and then loads the syringe. They have excellent results so far. The cultures have not stalled their growth and are shaken a few times daily.

[chill]

Just had a thought.

It was on my mind why people who pc their LC jars have success and yet boiling my water resulted in contams and no growth.

If the jar (or syringe in 'freaks case) are closed then maybe the air doesn't get boiled out entirely due to pressure but an open pan would dissipate all the O2.