Liquid culture trouble

[buster]

ay, tried to start a lc yesterday with some stem tissue and today there is just heaps of little brown specs in the lc ? this happened last time i tried, was just wondering if its a contam and i should toss it and start a new one ?

[myco]

if your going to do a tissue culture i would go straight to agar instead of a liquid culture

[Bigred]

i think the brown bits are the tissue of the mushroom decomposing,agar is the best way to go i can send you some if you need it

[buster]

sweet cheers for the replies, ye i think its screwed up might just toss it, it wasnt wild just one i grew.

Agar seems like the best option ay, cheers bigred but the little asian supermarket down the road sells it cheap as. bless them.

[BentoSpawn]

id definitely toss it !

spores>agar>LC

OR

tissue>agar>LC both work

Probably the best way would be

tissue/spores>agar>grain then squirt water into the grain and suck it back up into the needle

[buster]

is that once its fully colonized you do that bento? does there need to be any mycelium in the water you suck back up. sorry for noob questions last few attempts have been absolute shockers ha! so need all the advice i can get

[Bigred]

thats the best way to make a lc bento cant beat it

[Bigred]

it has to be fully collinized then get a sterile needle and water and squirt it in there you only need about 5 ml and thats enough to inoculate 12 jars

[BentoSpawn]

The advantage of this is you know the culture should be clean, its difficult to know your culutre is clean of you just pur spores in honey/karo water

[buster]

well that will work sick cause there is 2 colonized jars in the fridge, probably give it a go this arvo. cheers everyone for ya help.

[Bigred]

in a sterile environment fill a syringe with distiled water and put a very large needle on it place in pressure cooker at 15 psi for 15 min

allow to cool then shake up your jar and put it in a glove box or flow hood open the jar and squirt a little water on clean mycelium

and quickly suck it up thats about it your syringe is ready and you know its clean inoculated other jars but is it brf/verm or is it

grain as a grain to grain transfer may be your best bet

[buster]

aww yep i get ya, sweet this will be heaps better than makin syringes from spores every time ha! its grain so a g2g could be a possibilty but really want to start an lc so i reckon ill stick with squirting water into the jar.

[Bigred]

i find lc methods bring more vector;s (entry of contaminants) to the tek and grain to grain rocks trust me but each to there own .

i recommenced only using a little water as its ok to dilute mycelium and can last a very long time if you use distilled water

as the mycelium goes into hibernation . I would totaly recommenced looking into g2g (grain to grain) as once you have

a clean batch its theoretically can be done infinite times .

peace big red

[Psylo]

Red, and others, how many generations have you pushed doing G2G ? I'm up to G5 with some oysters, I figure I shoudl revert back to my original culture bank and start over. My understanding is that it's not 'infinate' because the biological ageing (senescence) will result in poor growth.

[goneski]

The next time I try LC, it will be cloning a tissue sample..I'll probably use honey again.

A couple of WBS quart jars I inoculated on 3rd with honey LC are around 90-95% colonised.. So insanely quick.

[Bigred]

im personaly on my fourth grg with portabelo's as long as you change the media ie barley then rye grain i dont think it will be a problem

i have noticed a jar that is about 2 months old has heaps of urine at the bottom how would one overcome this

[goneski]

Don't pee in your empty quart jars when drunk?

I thought 'myc piss' was a misnomer -- it's actually just metabolites and shouldn't really cause much of a problem?