Tissue Culture FAQ

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goldslinger

 
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Re: Tissue Culture FAQ

by goldslinger » Wed Sep 01, 2010 5:48 pm

Buy 2 plastic tubs. One to store Your prepared jars in, and a much larger one that You can turn upside down and cut out a square opening in the side for culture transfers, etc. That one would be Your 'clean room.' Some People use a fish tank turned on it's side as well.

Carol at Kitchencultures.com has good start up kits complete with manual and I think a CD.
Last edited by goldslinger on Thu Sep 02, 2010 2:22 am, edited 1 time in total.
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Re: Tissue Culture FAQ

by Hayden » Wed Sep 01, 2010 6:13 pm

Thanks, ill get looking on eBay for a few fish tanks!

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Re: Tissue Culture FAQ

by Hayden » Wed Sep 01, 2010 7:35 pm

Ive also just watched the video, he boils his media with the agar in the main mix. I get that you only add the agar in the pressure cooker stages, but do I have to boil the current mix?

Thankyou

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Re: Tissue Culture FAQ

by goldslinger » Fri Sep 03, 2010 12:27 am

All ingredients should be in the media before You boil it; sugar, Agar, MS, Vitamins, PGR'S (if You want) etc.

Agar type gelling agent isn't water soluable at all until boiled and requires cooking before dispensing to obtain uniformity of hardness or strength of Gelling properties from vessel to vessel. That is why People cook the prepared media. So technically You are cooking Your prepared media twice; once to dissolve the agar, and again in a pressure cooker, microwave, etc. for sterilization of the prepared media and the vessels holding it. This method gives You flexibility in how much media You want to dispense from vessel to vessel, knowing it will set up uniformly, irregardless of the amount. Agar gels upon cooling undisturbed.

You can get around the above problem of cooking twice by using a level 'smidgeon' measuring scoop per 25ml of media and dispense that little measure right into each of the vessels, swirl a bit and go into the autoclave which sterilizes and boils and dissolves the Agar all in one. You have to make sure You dispense the right amount of agar for the quantity of media in the vessel or You will end up too hard or soft on the media gel.

Carageenan type gelling agent, (I.E. GP-812) is quite popular as it is partially water soluable, dispersing uniformly so You can dispense into the vessels cold and only have to cook once (sterilization process) to activate it's gelling properties upon cooling.

With Carageenan type Gels, it's a good idea to occasionally stir the uncooked media while dispensing in the vessels, as the Carageenan can start to settle a bit making the first jars set up a little soft, and the last jars set up a little too hard.

No matter what Gelling agent You use, a little variance in hardness in media from vessel to vessel can be expected and is acceptable.

Hope this helps,

Gary
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Re: Tissue Culture FAQ

by Doomsday » Thu Jan 20, 2011 3:01 am

I am NOT an expert or even a decent...spert in any way at all, but this is what you should get imo.

A ten-20 gallon fish tank for doing the tc in as your sterile zone.
Some lights to grow out the jars.
A pressure cooker.
Baby food jars.
Autoclaveable caps
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Re: Tissue Culture FAQ

by LSK » Sun Jan 30, 2011 8:14 pm

1 "smidgen" spoonful it's = to how much grams of agar?

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Re: Tissue Culture FAQ

by Doomsday » Sun Jan 30, 2011 11:25 pm

If nobody here answers you first LSK I will measure it out on my gram scale tonight and pm you and post it here, but it is very little.
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Re: Tissue Culture FAQ

by Noddy » Wed Mar 16, 2011 9:18 pm

Can I ask a silly question here please

Is there a best light level, I know low lighting but the actual level that's best and duration in addition to that the Temperature that is also best for the plants...

Many thanks Noddy

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Re: Tissue Culture FAQ

by Matt » Wed Mar 16, 2011 11:13 pm

Noddy wrote:Is there a best light level, I know low lighting but the actual level that's best and duration in addition to that the Temperature that is also best for the plants...

I have read a number of foot candles before, but I don't recall what it was. Even though they grow OK with less light, I use the same amount of light for my cultures as I do for my plants in vivo that I grow under artificial lights. I think that Venus fly traps do better with stronger light even in vitro.

As for temperature, anywhere between 72°F and 82°F is optimal. Any warmer than that and you can cook them. Cooler than that and they don't grow as quickly.
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Re: Tissue Culture FAQ

by winterkid1 » Fri Mar 18, 2011 3:58 am

hey matt .........................
do you do tissue culture at your nursery??????????
and if you do ...can you send it to malaysia?????
i am looking for rare vfts that i cant find in malaysia.................
looking for : dentate,fused tooth,wacky traps, bristle tooth and other intresting vfts....................
hope you can TC them and ship to malaysia..................^^
thx,
sam

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Matt
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Re: Tissue Culture FAQ

by Matt » Fri Mar 18, 2011 5:18 pm

winterkid1 wrote:hey matt .........................
do you do tissue culture at your nursery??????????
and if you do ...can you send it to malaysia?????
i am looking for rare vfts that i cant find in malaysia.................
looking for : dentate,fused tooth,wacky traps, bristle tooth and other intresting vfts....................
hope you can TC them and ship to malaysia..................^^
thx,
sam

Hi Sam,

Yes, I do tissue culture and I can ship to Malaysia or anywhere else in the world. You can purchase sterile cultures here:
http://www.flytrapcare.com/store/intern ... onaea.html

Remember that a Phytosanitary Certificate is required for EVERY shipment. Also, Malaysia requires you to obtain an import permit. If you don't know how to contact the Malaysian government, I can help you with that. You can find their contact information here:
http://www.flytrapcare.com/store/about-sterile-cultures
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Re: Tissue Culture FAQ

by Dionae » Fri Mar 18, 2011 5:34 pm

Will TCs ever be available to US customers?

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Re: Tissue Culture FAQ

by jht-union » Fri Mar 18, 2011 7:35 pm

Dionae wrote:Will TCs ever be available to US customers?

I was wondering that too, the FTS international have more VFT's in TC for sale than plants available invivo for the U.S.A(so people not living in the U.S.A, consider yourself very lucky! ;) )
Can't wait when those are in stock for U.S.A people!
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Re: Tissue Culture FAQ

by Matt » Sat Mar 19, 2011 12:57 am

Dionae wrote:Will TCs ever be available to US customers?

Maybe, maybe not. I haven't decided yet.
jht-union wrote:I was wondering that too, the FTS international have more VFT's in TC for sale than plants available invivo for the U.S.A

That's because it takes a long time to plant out the tissue and grow it to a reasonable size for sale. Everything that's in stock in the FTS international store in vitro will eventually be for sale within the US in vivo (planted out).
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Re: Tissue Culture FAQ

by BradR » Sat Mar 19, 2011 3:45 am

Matt wrote:
Dionae wrote:Will TCs ever be available to US customers?

Maybe, maybe not. I haven't decided yet.


Don't do it Matt. You worked too hard getting all those clones into TC to hand them over to your competitors.

Just my two cents worth.
Brad
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