Store bought Liquid Culture Medium...

[johnnydigital]

I had read something in the archives about store bought liquid culture mediums. Someone had the wonderful idea of injecting spores into a Vitamin Water bottle... and it worked!

Well, I had the same idea and a couple of weeks ago, I started my experiment. I chose an electrolite recovery drink called 'Pedialight' for infants as my Liquid Culter medium. The only reason I chose this was because I had a 6 pack of it from a sick kitten we were trying to nurse back to health. This was 4 years ago and the bottles were even expired.

A tissue sample was taken of the target mushroom, dunked in Hydrogen Peroxide for 5 seconds, and then dropped directly into the Pedialight solution... as is. No modifications were made to the bottle and everything was done open air. The solution was shaken roughly and the lid screwed on, but left very loose. I also did the same procedures to a 50/50 solution of Pedialight and H20, and the same thing with a typical Karo tek. So, 3 LC's total. 1 pure Pedialight, 1 50/50 Pedialight and 1 Karo tek. The LC's were incubated in the normal fashion and were roughly shaken every 4 days.

Well it's been 2 weeks and the results are in... the Pure Pedialight LC is the clear winner! The growth is tremendous, the solution is almost totally cloudy. The 50/50 Pedialight and the Karo Tek jars are OK, but not nearly as colonized as the pure Pedialight.

I think this will prove to be very helpful to folks who want to get into cloning and LC's. It really couldn't be easier, as no preparation is really needed. Just go get a bottle of Pedialight, throw a tissue sample in it and incubate. In 2 weeks you'll have enough solution to innoculate 500 jars. It's just that easy.

I'll try to take a pic, but LC's usually don't show too well in pictures. Besides, all I got is my crappy camera phone.

[LotRev]

Wow, that sounds pretty good. Nice and Easy.

Could you possibly post some pictures of the LC's?

[johnnydigital]

Ya, I will post pics of the LC once I can get my hands on a decent camera. I doubt any of the myc will show up in a camera phone pic. Here is the Pedialight I used tho.

[Hippie3]

have you actually tested it yet to be sure it's clean mycellia in there ?
seems open-air lid-off transfer would increase contam risks considerably,
when i did this
i put a silicon port on the un-opened bottle
to inject thru,
ensuring sterility.

[johnnydigital]

Oh, no doubt its myc... but just started the testing to be sure. A small sample was taken and placed in agar...so, we'll see whats up in a couple days. SWINJD works in a clean, bleached environment and I work quickly so it shouldnt be an issue...

Also, the little bottles aren't quite like the one SWINH3 used, in that, they have one of those tamper proof lids under the cap... know what I'm talking about? It was peeled up halfway and the extracted tissue sample was then placed in the solution. It acts sort of like a little hood to prevent shit from falling in the bottle.

[trips6928]

Lookin forward to seeing you results. I have some Parent's Choice elecrtolyte, unflavored and fruit flavored. I added a myc strand to both, and I can clearly see the strands growing after 2 days(very ropey looking). The fruit flavored one seems to be quicker probably cuz it has extra ingredients.
Anywho...good luck with ur tests.

[Coopdog]

Thank you for that idea! swim was just battling with the idea of using one of the last three sporeprints to do a liquid culture, and you just made it just the way he like it, easy as hell Peace....

[valleyofmushies]

i will try this , thank you...i need to speed up LC time ...its a month right now
this is another example ..i love(care) about this place

[johnnydigital]

Here is a shitty pic of the LC bottle. This is the 100%, unaltered store bought 'Liquilytes' bottle. Jar was opened, tissue was placed in the jar, it was shaken and incubated... this pic is from the 10th day of incubation and the growth is profound. This pic doesn't really do it justice, but you can clearly see a mass of growth at the bottom of the jar even with this shitty pic. In real life, the bottle is extremely colonized... much more than any LC SWINJD has ever done.

Now for the next step.....LC solution was drawn and placed on 2 PDA agar petri dishes, then 1 PFTek BRF jar was also nocced up with the LC solution.... More updates to follow.

[zodd]

oww ill be checking this out in future.

good luck johnny.

[coconut1225]

that is absolutely amazing. This idea could cut a major corner of the growing process without risking too much time and effort. Sounds great. Im gonna try this method with a store bought white button for my wifes first grow

[BlazedChillin]

I'm with hippie I wouldnt feel safe opening up in open air, but I may give it a shot in the glove box.

I can see the clump in the bottom, but it also looks cloudy....could be contam, could be the pic. Cant wait to see what the agar looks like.

[johnnydigital]

When I said 'open air' I suppose I was a bit vague. The procedure was done under clean conditions, just not in a glove box. The lid of the bottle was never fully removed during the process and the sample went from the peroxide bath to the 'drink' within about 2 seconds.

The 'drink' contains a few goodies that the Karo Tek does not.. including Sodium Chloride and Tartaric Acid which help to inhibit bacterial growth. I guess that's why when you leave them open, you don't get a bunch of gunky shit in there right away. The solution is salty, but sweetened with stuff that's 200x more sweet than sugar, to mask the saltiness.

[Getitosis]

just come across this.

interesting jd...

how's that agar looking?

[johnnydigital]

Little update.

Samples of the LC were taken and transferred to agar. Also, a syringe of LC was made and injected into a simple 1/2 pint PF jar. Well, the agar is looking fine... no issues there. But, the LC that was injected into the PF jar turned out green.

I think the problem was I used an old syringe to knock up the PF jar (one that was contaminated when I got it from a vendor) and it wasn't cleaned properly. I just tried the same procedure using a fresh, sterile syringe, so we'll see what happens. Long story short, there's definitely healthy myc growing in the LC. Not bad from a 1/4" x 1/8" piece of starter tissue. I have many more bottles of this Pedialyte stuff, so hopefully this process will work.

I did add a polyfill filter to the cap of the bottle for gas exchange and that seemed to help quite a bit. Next I'm going to add a silicon 'port' for making LC syringes.

Overall, I think it's a good source for Liquid Cultures, as it needs NO pressure cooking or sterilization and only minimal (and easy) modifications to the actual store bought bottle.

[Azuloso83]

Good stuff johnny. Just a couple questions....

Do you think this would work for a multispore injection?

Would it be ok to transfer the pedialite to a jar that already has a modified lid for ease of use?

If multispore is ok, how much is necessary for say one bottle of pedialite?

Thanks and good luck!

[Hippie3]

the LC that was injected into the PF jar turned out green.

bummer.

[Hippie3]

Originally Posted by Azuloso83

Good stuff johnny. Just a couple questions....

Do you think this would work for a multispore injection?

Would it be ok to transfer the pedialite to a jar that already has a modified lid for ease of use?

If multispore is ok, how much is necessary for say one bottle of pedialite?

Thanks and good luck!

sure multispore works,
assuming decent concentrations
1 cc should be plenty per bottle.

[zodd]

the fact that youve got growth is more than a success. even if the pedialite dont come sterile, (which is unlikely) you would still save a tone of time compared to boiling draining, straining and eating potatoes before the sterilizing.
just pop that in a microwaveable container and take a second stab at it.
rooting for you jonny.

[Azuloso83]

Thx Hip, my FOAF will see if he can replicate johnny's apparent success.

[johnnydigital]

OK, another update here. The syringe was def contamned. I went out and bought new, sterile, clean syringes on friday and re innoculated 2 PF test jars. After 2 days of incubation, they show signs of strong growth.

Overall, i think it was a success, but few points...

1. First, use sterile syringes!!!! If you have a syringe that has been contaminated, either chuck it, or PC it.
2. Second, the initial transfer of the clone sample into the LC should be done in the most sterile of conditions. I did 3 LC jars, open air, and only 1 took.... Huge waste of space and materials by the time you figure in the incubation of the LC, the test jars, etc.
3. Third, gas exchange is necessary. So, modify the lid to allow for gas exchange. Also, if one were so inclined, one could add a dab of silicon to act as a self sealing port for the withdrawl of LC. Both of these are great ideas and they really cut down on the risk of contamination.

Overall, I think there is room for store bought liquid culture medium. If the proper care is taken, one can get a whole lot of LC from small, store bought bottles.

[johnnydigital]

One other thing, if one wanted to transfer the contents of a store bought LC to another bottle, then I would PC it. The point of it all is to attempt to obtain a strong LC with minimum steps. But, my results thus far show that pedialyte works for LC, so transferring it and PC'ing it should work fine.

[samspad]

Originally Posted by johnnydigital

Little update.

I did add a polyfill filter to the cap of the bottle for gas exchange and that seemed to help quite a bit. Next I'm going to add a silicon 'port' for making LC syringes.

Overall, I think it's a good source for Liquid Cultures, as it needs NO pressure cooking or sterilization and only minimal (and easy) modifications to the actual store bought bottle.

If you make those modifications to the store bought bottle, wouldn't it have to be PC'd?