Storing Liquid Culture
|Posted by: Peroxide Feb 11 03, 06:00 AM GMT|
| Ok so I have tried to explain this a few times. Here is my theory, and some info to back it up.
the basic concept is to combine the functions of a syringe and a honey jar. Useing peroxide to destroy possible contams.
so after finding a good isolate, get some of its myc into a syringe. That is 1/2 full of 3% peroxide diluted to 6ml/liter. Once youve got the myc in the syringe fill the syringe the rest of the way with the usual honey tek mix treated with peroxide in the previously mentioned ratio.
Let this incubate, and then use it to knock up a jar and grow from there. I figure you can easily get 10 jars a syringe. before you have to feed the myc again.
The syringe should be stored in the fridge before all nutes are used. when using the syringe leave the last cc. This is your starter.
My calculations for the useful life of the syringe are based on these assumptions.
Ageing (senescence) is caused by total number of cell divisions (stamets)
senescence is approximately 1/3 as apperent as in a pint jar (forums) anno agrees.
Senescence sets in around 3 generations of transfers (lowest numbers on forums)
Myc in a liquid is still viable for 2 months in the fridge.
You should be able to resample tissue from the isolate at least twice. before going back to spores or stock.
Now the math. 3 generations at 1/3 weight * 2 months for the fridge * 3 total isolations (3 / 1/3)*2*3 = 54 months.
Contams; judicious use of peroxide and having multiple syringes of the isolate running at the same time would be a good idea. with 4 syringes, I am sure you could figure out a rotation to ensure enough viable myc even if 2 contam to run the full lifespan. While significantly slowing the ageing.
Anyway the reason for this post since I have no hard data to back this up is to ask did I miss something or is this really going to work?
|Posted by: OneDiaDem Feb 11 03, 06:58 AM GMT|
|My liquid innoculant stayed viable for 10 months at last drawing. Same colonization rate, same fruiting characteristics. I dont see why this wouldne be something to have a go at.|
|Posted by: DirtyWOP Feb 11 03, 09:46 AM GMT|
| Why not just use one large stock culture in the fridge, and add honey water/draw syringes as needed....so you don't have to keep it in a syringe.....
And using that much pure peroxide is probably too much....
|Posted by: Fungusmaximus Feb 11 03, 04:32 PM GMT|
|When storing cultures for extended periods its a good idea to have a wide range of nutes for the myc to consume, this will aid in keeping the myc's vigor and speed and keep its defense against contams up. Too much peroxide for an extended period will retard and mutate growth if using H2o2, its best to give it a quick bath in the h2o2 and then refill with nute solution. I have poured straight h2o2 into badly contamed clone cultures for 3 days + and effectively killed all contam spores, but when used as inoculant the myc grew at a very slow snail like pace. Where as before it was colonizing pints of WBS in 11 days...|