Plant Tissue Culture Question

Hello! I have been reading up on how to do plant tissue culture and today is the day I'm finally going to do it and I just realized that there is something that I am unclear about and I can't find the answer! Once I have sterilized the medium, cooled it and put my plant cuttings into the jars of media, am I supposed to put the covers on completely tight or leave them loose? I've read that you wrap the lidded jars in plastic wrap after covering them, so I'm wondering if you have to do that because the covers are left loose. Could you tell me? Tight, or loose? Thanks a lot!!
Deb

i tried tissue cultures just last month. it didnt pan out bc i was unprepared to battle the heat we had so they didnt survive stage three. but you can put them on tight or leave them loose. the main purpose of the plastic is to keep mold spores out. just dont blow air directly on the vessels, it increases your chances of contamination. i hope this helped and good luck. finding any information on tissue culturing is pretty tough.

al lot of people use tyvek and rtv sealant to make filters other wise keep thos lids on tight. they will probably not mature enough to need more air than the vessel can provide for a while.

honestly ive dabbled in TC and mycology, the rate of maturation with TC makes it very difficult to preform without a HEPA filtration system, otherwise youll end up with a lot of cotaminated vessels becauese of all the switching needed to get cultures to a plantable point.

if your really serios about TC i would look into how to build hepa filtration, and buying a tent for the purpose specifically thats what im doing, i bought three tents to grow in for now since my 5x5 is too big for the area right now but once i start using that again i will use one of the three tents as a TC culture grow/work area.

also if building a room for your TC your HEPA filtration system will need to be at least twice as many CFM(after the 20-30% for filter reduction) for the area, to create positive pressure to keep out unsterilized air.

"and this has been flips quick tip"

I have never seen any reason to use TC for herb. I do use it for hard to propagate carnivorous plants. If you really want to give it a shot you will have to look into the correct protocol for the plant. Maybe one for a tomato or a pepper plant? You can get special vessels that are designed for TC but a lot of people use baby food jars with special lids. Then wrap the lids up tight with plastic wrap that's been cut to size while still on part of the tube. Most of the agar media that's used has anti-biotics in it but if you botch the batch you still run the risk of getting contaminates growing in with the plants.

I use a still air cabinet that has been washed out with bleach and I make sure everything that is used is sterilized VIA pressure cooker. If you wanna make a laminar flow hood there are some good designed that incorporate HEPA and UV filters that can be built for a little bit of money but not for a few hundred because the correct HEPA filters aren't cheap.

Here is where I get my media, Hormones and PGR's from.
http://www.hometissueculture.org/
I use to buy from Sigma-Aldrich but the above link is way way cheaper but here is the link to the protocols that Sigma provides. Ill have to look more for the plant specific protocols that sigma sent to me as a book. Hopefully they have a digital version.
http://www.sigmaaldrich.com/life-science/molecular-biology/plant-biotechnology/tissue-culture-protocols.html
In fact here is the literature request. The book you want is the "Plant Tissue Culture" under life sciences section. They should send it to you for free. If they don't let me know and Ill copy a few protocols from the book or I might be able to scan it into a PDF format for you.

Also here is a youtube page that gives some really good info on how to do TC at home. He knows his shit and breaks it down to make it easy.
http://www.youtube.com/user/fbt2007

Good luck and I wish you the best. TC is fun but it can be frustrating till you get a good handle on the process.

There is a reason for it but like you've all point it out it's very hard to do. I heard of people trying for over a year before they got it to work for them. Lots of trials and errors. Many ways to do it wrong.

But, if you're a strain whore and can never get rid of a strain or are a breeder that wants to keep a back up of genetics then TC is awesome. But like people have said.. steep learning curve and good equipment makes a big difference.

I know Bill Graham from http://planttc.com/ and have talked about it a few times with him.

I built my shroom culture cab from Sterilite and put HEPA filters on it. Bought HEPA filter replacements for vacuums and simply Gorilla Glued them over holes I cut with a hole saw. Works great so far.

eDude said:

There is a reason for it but like you've all point it out it's very hard to do. I heard of people trying for over a year before they got it to work for them. Lots of trials and errors. Many ways to do it wrong.

But, if you're a strain whore and can never get rid of a strain or are a breeder that wants to keep a back up of genetics then TC is awesome. But like people have said.. steep learning curve and good equipment makes a big difference.

I know Bill Graham from http://planttc.com/ and have talked about it a few times with him.

Click to expand...

Thanks eDude. We are always trying to keep it simple and successful. You may be interested in one of our simplified methods set up after so much travel and TC presentations...liquid tissue culture starts. Here is a short write-up and pics. Good luck.