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Tissue Culturing & Other Propagation Techniques of Banana Plants This forum is for discussing propagation techniques of banana plants. Tissue culturing is the popular process of creating clones from a source plant. There are other techniques to propagate banana plants however, such as nicking corms or dividing corms. Learn more inside.


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Old 09-16-2009, 12:10 PM   #1 (permalink)
 
Location: CAracas, Venezuela, South America
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Default Almost ready for ordering...just a few doubts

IŽm about to place an order but I still have a few doubts:

Is BA the same as BAP?
I canŽt understand why Caisson states the following:

6-Benzylaminopurine (6-BA, BA, BAP)


IŽll be using baby food jars ( the slightly taller ones ) with metal caps.
Would I have to drill a hole for gas exchange?

IŽll be using the following products in POWDERED form:

Kinetin, IAA, IBA and NAA. Some paper states that I can dissolve these in
Isopropyl alcohol.
BAP can be dissolved in NaOH. I can get lye whose composition is as follows:
NaOH 95%, NaCO3 1% and inert ingredients 4%.
Is this OK: is there anything else I can use?

I am particularly interested in plantains and poinsettia. Of course IŽll try avocado, mango and cashew.
Some protocols call for media with inositol and thiamine ( Keith Benson )
Do you recommend MS Basal Salts ( M524 ) or should I order MS with MS vitamins? Thanks, everybody.
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Old 09-16-2009, 05:46 PM   #2 (permalink)
 
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Default Re: Almost ready for ordering...just a few doubts

Quote:
Originally Posted by alah View Post
Is BA the same as BAP?
Yes.

Quote:
Originally Posted by alah View Post
I canŽt understand why Caisson states the following:

6-Benzylaminopurine (6-BA, BA, BAP)
3 abbreviated synonyms.
Benzyl Amino Purine
BAP

Quote:
Originally Posted by alah View Post
IŽll be using baby food jars ( the slightly taller ones ) with metal caps.
Would I have to drill a hole for gas exchange?
Yes. Then put a filter over it so contaminants don't get in.
Or use Magenta Caps.

Quote:
Originally Posted by alah View Post
IŽll be using the following products in POWDERED form:

Kinetin, IAA, IBA and NAA. Some paper states that I can dissolve these in
Isopropyl alcohol.
BAP can be dissolved in NaOH. I can get lye whose composition is as follows:
NaOH 95%, NaCO3 1% and inert ingredients 4%.
Is this OK:
It should work.

Quote:
Originally Posted by alah View Post
is there anything else I can use?
You can use NaOH.

If you are trying to do TC in a non-lab setting, and/or with limited access to supplies, I recommend you go visit

Hometissueculture.org (formerly Kitchen Culture Kits - Intro Page )

for a while, and join the forums, before you spend your money. They can show you what is a good substitute, and when a substitute isn't a good idea.

Adrift ( KJ, Ken, Kenny, bonehead )
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Old 09-16-2009, 09:48 PM   #3 (permalink)
 
Location: CAracas, Venezuela, South America
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Default Re: Almost ready for ordering...just a few doubts

Quote:
Originally Posted by adrift View Post
Yes.



3 abbreviated synonyms.
Benzyl Amino Purine
BAP



Yes. Then put a filter over it so contaminants don't get in.
Or use Magenta Caps.



It should work.



You can use NaOH.

If you are trying to do TC in a non-lab setting, and/or with limited access to supplies, I recommend you go visit

Hometissueculture.org (formerly Kitchen Culture Kits - Intro Page )

for a while, and join the forums, before you spend your money. They can show you what is a good substitute, and when a substitute isn't a good idea.

Adrift ( KJ, Ken, Kenny, bonehead )
Thanks, KJ. I joined the the forum about 3 months ago. Since then IŽve been to India, Pakistan, the US, ALL over this forum, THTC, and a few other places I donŽt even know. I decided to order powder because it very much cheaper and weighs much less ( otherwise shipping would be prohibitive );
Due to this, I plan to order CaissonŽs minimums which would seem a lot for many of you but I canŽt run out of supplies when I finally get to the midddle of this. IŽll wait till Carol comes back, anway. Thanks a lot.
Off Topic: If any of you are from South Africa or nearby, can you please contact me, Alberto.
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Old 09-17-2009, 04:23 AM   #4 (permalink)
 
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Default Re: Almost ready for ordering...just a few doubts

Dear Alberto,

in my experience the best growth stimulators or hormones, are the ones that are heat resistant. IBA, NAA, BAP, Kinetin are heat resistant others such as GA3 (gibberelic acid) or TDZ (thidiazuron) are not autoclavable and have to be added to media after autoclaving using a filter to cold sterilize them. Keep this in mind. Also you can use household caustic soda (little granules) to prepare a 1N Naoh solution (40 g of NaOH dissolved in distilled water and made upto 1 litre gives 1N NaOH solution)
measure 25mg of the hormone and disolve in 1N naoh (as little as possible) then add up with distilled water until 25ml, this will give you 25ml containing 1mg of hormone per 1 ml of water. you can store these in the freezer for a long time. do not use undiluted naoh in your media you will have to seriously adjust ph if you do. ( the product of neutralising NaOH with HCl is NaCl or salt the more you neutralise the salter your media, not good that is!)
i have for you, if you wish, protocols for micropropagating mango cashew and avacado

good luck my friend
any questions? ask!

Dave

ps. if you dissolve hormones in alcohol they may come out of solution when you add them to water a better solvent would be DMSO (dimethylsulfoxide) even better then naoh.
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Last edited by rohsen : 09-17-2009 at 04:38 AM.
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Old 09-17-2009, 03:58 PM   #5 (permalink)
 
Location: CAracas, Venezuela, South America
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Name: Alberto Albert
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Default Re: Almost ready for ordering...just a few doubts

Quote:
Originally Posted by rohsen View Post
Dear Alberto,

in my experience the best growth stimulators or hormones, are the ones that are heat resistant. IBA, NAA, BAP, Kinetin are heat resistant others such as GA3 (gibberelic acid) or TDZ (thidiazuron) are not autoclavable and have to be added to media after autoclaving using a filter to cold sterilize them. Keep this in mind. Also you can use household caustic soda (little granules) to prepare a 1N Naoh solution (40 g of NaOH dissolved in distilled water and made upto 1 litre gives 1N NaOH solution)
measure 25mg of the hormone and disolve in 1N naoh (as little as possible) then add up with distilled water until 25ml, this will give you 25ml containing 1mg of hormone per 1 ml of water. you can store these in the freezer for a long time. do not use undiluted naoh in your media you will have to seriously adjust ph if you do. ( the product of neutralising NaOH with HCl is NaCl or salt the more you neutralise the salter your media, not good that is!)
i have for you, if you wish, protocols for micropropagating mango cashew and avacado

good luck my friend
any questions? ask!

Dave

ps. if you dissolve hormones in alcohol they may come out of solution when you add them to water a better solvent would be DMSO (dimethylsulfoxide) even better then naoh.
Thanks a lot, Dave. I got that straight.
These PGRs can be dissolved with 1Normal Sodium Hydroxide. Just place the weighed amount in a glass vessel and add the solution, maybe drop by drop while stirring, until it dissolves. Add distilled water up to the required volume which depends on the weight of the PGR. Of course I should be able to use less water and so have a stock solution, maybe X10 or X20.
As for those protocols, they really interest me. Please let me know how to proceed. One other point, I rermember your idea about posting info Levels 1, 2 and 3. IŽm working on a Level 1, of course, which IŽll be posting soon.
Again thanks... to everybody.
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