My 1st. TC experience

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Highlander

 
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My 1st. TC experience

by Highlander » Thu May 09, 2013 2:04 am

Hey guys!

I want to try a simple experiece with my sundews and my vft.

I´ll describe my equipments and supplies below:

Equipments

- autoclave 23liters
- magnetic stirrer
- digital phmetro
- digital ppmmetro
- pipettor auto(0,5ml)
- water distiller
- digital scale 0,05g precision
- laminar flow hood( my own project)

Supplies

- PPM
- MS powder from planttc as multiplication labeled
- MS powder from planttc as stabilish labeled
- NAA
- IBA
- pure agar
- KOH (ph + )
- HPO4 (ph -)
- food coloring
- alcohol ethanol 70%
- alcohol ethanol gel 46%
- alcohol isopropyl
- tween 20
- H2O2 6%
- captran(fungicide)
- bleach 10%
- bleach 5%
- bleach 3%
- bleach 1%

***********************************************

1 - Can I start something or I need to buy some supply/equipment?
2 - What kind of sugar I have to use? Here(Brazil) we have 4 or more types, can I see a photo of the correct sugar?

Thanks :)

Best regards
Ricardo
Last edited by Highlander on Tue Jul 02, 2013 10:50 pm, edited 1 time in total.

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coline

 
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Re: My 1st. TC experience

by coline » Thu May 09, 2013 2:29 am

Way to go, you're nicely equiped!
Well, first of all, as you may have seen in the PPM bottle, just as a precaution, you may not use the Tween 20 with the PPM when working, it disables the effect.
Then, as a sundew proyect, you may be able to start as the drosera may use a plain MS medium. But for other plants or achieving different results in multiplication or rooting you will need some plant growth regulators as BAP, NAA, Kinetin, AIB or AG3, te first 3 are the most used.
Also, what exactly does that MS medium you have contain?
The sugar is simple, the cheap white sugar anyone uses to add to coffee or drinks is the sugar you want.

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Highlander

 
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Re: My 1st. TC experience

by Highlander » Thu May 09, 2013 3:07 am

Hello Coline, thanks for help!

I forgot to say that I have baby food jars...teste tubes....all autoclavable products.
I also have GA3!

I don´t know what my ms contains...because the bottle don´t show any info...only the label multiplication or stabilishment/roots.
Here we have sugar very white(called refined sugar) and another used to make cakes...another called(maskavo).....so I´m afraid I´m using wrong sugar :(

Best regards
Ricardo

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coline

 
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Re: My 1st. TC experience

by coline » Thu May 09, 2013 3:14 am

The sugar, the one used for cakes should be fine, maybe you got to know here the brands Juan Viñas or Doña María, those kind of normal sugar, the powder is expensive but it is also possible to use.

That you do not know what the MS media contains, and as it says that it is MS specifically to do something makes me think it already has some regulators inside, which normally may not be in the right concentration for your plants.
The best is to get to make your own MS or buy a premixed version, but with nothing more than macro&micro elements and vitamins. Places as kitchenculturekits, caisson labs, phytotech are good sources for the medium and any other supply.
You have everything needed to make the TC start, just the MS is in doubt, try to contact who sold it to you.

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cvlover

 
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Re: My 1st. TC experience

by cvlover » Thu May 09, 2013 4:24 am

You can start TC with your list equipments. Some tips:
- Sugar: white food-grade sugar is good enough.
- It's better to get some different popular medium: MS, DKW, WPM, ...
- Get cheap and well known powder hormone: BA, NAA so you can make medium with different hormone concentration.
- Remember, TC doesn't stop at VFTs and sundews. You can play with orchids, herbs, woody plants aquarium plants and mushroom etc...

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Highlander

 
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Re: My 1st. TC experience

by Highlander » Thu May 09, 2013 2:15 pm

coline wrote:The sugar, the one used for cakes should be fine, maybe you got to know here the brands Juan Viñas or Doña María, those kind of normal sugar, the powder is expensive but it is also possible to use.

That you do not know what the MS media contains, and as it says that it is MS specifically to do something makes me think it already has some regulators inside, which normally may not be in the right concentration for your plants.
The best is to get to make your own MS or buy a premixed version, but with nothing more than macro&micro elements and vitamins. Places as kitchenculturekits, caisson labs, phytotech are good sources for the medium and any other supply.
You have everything needed to make the TC start, just the MS is in doubt, try to contact who sold it to you.


Hey Coline, thanks one more time!

We have 4 types of sugar, see below:

Image

I used the white sugar with very small granules.

I´ll buy this itens:

Media MS - with vitamins?
hormones - BAP, NAA, Kinetin.

Thanks

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Highlander

 
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Re: My 1st. TC experience

by Highlander » Thu May 09, 2013 2:16 pm

cvlover wrote:You can start TC with your list equipments. Some tips:
- Sugar: white food-grade sugar is good enough.
- It's better to get some different popular medium: MS, DKW, WPM, ...
- Get cheap and well known powder hormone: BA, NAA so you can make medium with different hormone concentration.
- Remember, TC doesn't stop at VFTs and sundews. You can play with orchids, herbs, woody plants aquarium plants and mushroom etc...


Hey cvlover!

Thanks for the tips!

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coline

 
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Re: My 1st. TC experience

by coline » Thu May 09, 2013 2:23 pm

Ok, I see, well the white top type is the good one, it has low impurities or other compounds from the sugar cane that may alter the medium.
Here we have those, but we have even more types, powder white, fine white, coarse white, brown (would be the second sugar), dulce de tapa (third or 4th sugars), tapa de dulce, and molases. And not counting that fructose is sold also.

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Highlander

 
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Re: My 1st. TC experience

by Highlander » Fri May 31, 2013 4:46 pm

Hello Friends!

I just received this products:

Gellan Gum Powder:
Image


PPM:
Image

MS basal with vitamins:
Image

Mix for plant tc:
Image

Remember that I have:

Equipments

- autoclave 23liters
- magnetic stirrer
- digital phmetro
- digital ppmmetro
- pipettor auto(0,5ml)
- water distiller
- digital scale 0,05g precision
- laminar flow hood( my own project)

Supplies

- PPM
- MS powder from planttc as multiplication labeled
- MS powder from planttc as stabilish labeled
- pure agar
- KOH (ph + )
- HPO4 (ph -)
- food coloring
- alcohol ethanol 70%
- alcohol ethanol gel 46%
- alcohol isopropyl
- tween 20
- H2O2 6%
- captran(fungicide)
- bleach 10%
- bleach 5%
- bleach 3%
- bleach 1%

I bought also(still waiting to arrive) this products:

- Indole Butyric Acid
- Naphthalene Acetic Acid

Can I start with this tools/products or have to wait/buy others supplies?

Thanks in advance!

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coline

 
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Re: My 1st. TC experience

by coline » Fri May 31, 2013 6:04 pm

You may start correctly now

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cvlover

 
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Re: My 1st. TC experience

by cvlover » Fri May 31, 2013 6:47 pm

Highlander wrote:Hello Friends!
.....
I bought also(still waiting to arrive) this products:
- Indole Butyric Acid
- Naphthalene Acetic Acid
Can I start with this tools/products or have to wait/buy others supplies?
Thanks in advance!


You can start now without hormones. Sterilization the most important process in TC. Fail to do it you shouldn't talk about TC. Try seeds first. Choose quick and easy-germinate seeds so you will feel happy and not frustrate with contamination. Good luck my friend.

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coline

 
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Re: My 1st. TC experience

by coline » Fri May 31, 2013 6:59 pm

Very good seeds are Dionaea, they are very big and easy to dissinfect. also, as you live in brazil, it is really easy to get unoppened orchid capsules, you may try to introduce the seeds from them also.

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Re: My 1st. TC experience

by Matt » Fri May 31, 2013 7:13 pm

cvlover wrote:You can start now without hormones. Sterilization the most important process in TC. Fail to do it you shouldn't talk about TC. Try seeds first. Choose quick and easy-germinate seeds so you will feel happy and not frustrate with contamination. Good luck my friend.

Perfect advice!!!
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Highlander

 
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Re: My 1st. TC experience

by Highlander » Fri May 31, 2013 7:45 pm

Hey friends!

Thanks for help!

I just bought a tipical VFT and I´ll try to start my exp. using this plant.
I think to do this stepbystep:

Preparing the media:

1- Dissolve the MS+vit powder in 900mL of destilled water and stir
2- Add the mix for plant tc
3- Add 30g of suggar and stir
4- Add food coloring and stir
5- Add 2,5g of gellan gum powder(see picture) and stir
6- Complete with destilled water up to 1L mark
7- Get the ph of the solution and adjust until ph5.6

Autoclaving the media:

1- Put the media into a 1L autoclavable bottle
2- Put all test tubes+caps
3- Put all bottles with destilled water(for sterilization step)
4- Put scalpel/forceps...etc

After autoclaving dispose the hot media into the test tubes and cap.

I think do my 1st. exp. in 2 days, first day I make the media and sterilize water,tools etc. In the second day I sterilize the explants and put them into the media.

To sterilize my VFT explants, I wanna do this process:

1- wash the explant in current water about 2 minutes
2- wash the explant in destilled water about 2 minutes
3- wash the explant in destilled water+PPM about 4 minutes
4- wash the explant in destilled water about 2 minutes
5- wash the explant in destilled water about 2 minutes
6- Put the explant in a esterilized plate and cut off top(trap)/botton(white tissue) parts
7- Put the explant into the media and cap the tube using a plastic film as a seal.

I know that probably I made some mistakes in my step-by-step, so I need to learn about my mistakes using yours help

Many thanks to you all once again!

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rellenburg

 
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Re: My 1st. TC experience

by rellenburg » Fri May 31, 2013 9:16 pm

As for the autoclaving media section I would...
1. Pour prepared media into a pot (or other container that is safe for a stove top or hotplate)
2. Heat on stove or hotplate almost to a boil while stirring media until everything dissolves.
3. Dispense hot liquid (you can let it cool a little) into vessels and secure lids.
4. Autoclave at 15psi for 20-25 min (depends on how much media you're making)
5. When cooled and gelled it is ready for use.

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