Liquid culture cloning questions

[p2k1]

Hey everyone Im attempting to clone some outdoor fruits using liquid culture. Im looking to keep things as simple as possible. Im wondering if I can dip whole fruits in peroxide and then place them in the liquid culture? How about cutting out the middle segment of a stem? (The stem above the base and beneath the cap) Do I need to split the fruits and cut some tissue from the inside?
Thanks

[p2k1]

I dont have a flowhood or agar or even a syringe to do a biopsy. The plan is to clean a bathroom, disinfect heavily, stop all air flow. Im most likely going to clean a box, set it on its side and sterilize it so I can work in the open side. Plan is to microwave 4% honey liquid culture and place in box along with a shallow container of hydrogen peroxide, sterilize the air again and quickly either twist a fruit to expose tissue, collect some with a knife or skewer and dip it in peroxide and then immediately transfer into the liquid culture in the sterilized box. The other option would be to cut off the bottom and top of a stem and dip it in peroxide and then transfer it, unless it would be contaminated on the outside. They were grown outside so if the peroxide doesnt sterilize it ill probably have to use an inner tissue simple. Ill probably make atleast 2 LCs, one with a tissue sample if I can pull it off and the other with a stem chunk

[Boboso]

Sounds like a hell of a gamble.

Your best bet IMO would be to either PC an old syringe, or get some new ones first. That way, you can do a simple LC with less worry about contams.

You'd be able to wipe the side of the stem with ISO, then stab in with a syringe with a lil water, squirt, then suck some mush in and squirt that into the LC.

To make the LC simple, you could do 2 cups water, 1 tablespoon honey, 1 tablespoon karo, toss it in a jar with RTV sealed lid, and toss into the oven at 300 for an hour. IMO, this lazy oven tek is a great way to go, because you wont be opening a jar repeatedly, and the moment it comes out of the oven, everything is sterile inside.

Then again, I don't exactly use a glovebox or flowhood, so anything that involves opening a jar prior to birth makes me a little uncomfortable...

[BuckarooBanzai]

The main problem is that if the LC is contaminated, not only is it useless you won't know it is useless until you test it and contaminate other things.

The cool thing about agar is that you can very easily visually identify contaminants. Very hard to do that in liquid suspension.

Do you have access to any half pint jars? You might try my cloning Tek. It uses LC, but just a little so you can see the growth more clearly.

http://forums.mycotopia.net/cloning/...ek-merged.html (Buckaroo's Cloning Tek...{merged})

G'luck regardless!