My TC adventure

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tokol0sh

 
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My TC adventure

by tokol0sh » Wed Dec 12, 2018 5:08 am

Hi All,
I am looking at restarting my CP collection (between a new dog, chickens, university and a few years of neglect al my CP's died). I have also tried tissue culture at home before (growing potatoes) and that was reasonably successful. I figured I would try putting some CP's in tissue culture and repopulate my garden. I mainly interested in Dionaea, Drosera and Sarracenia varieties.

It has been a while since I last tried TC (~6 years) and am just about to place an order with PhytoTechnology. I am looking for some feedback on my order list (basically, have I overlooked anything). I have also added some growth regulators for experimentation, following this paper: http://sci-hub.tw/10.1023/A:1021203811457

The current order list:


I intend to dilute my MS media down to 1/2 or 1/3, I am a bit unsure on some terminology. Some recipes call for 1/3 MS with full vitamins, others say 1/3 MS with vitamins and others will just say 1/3 MS. If I buy MS with vitamins and dilute it 1/3, do i need to add more vitamins to bring it back to "full" vitamins? If so then would something like this be needed:
(M557) Murashige & Skoog Modified Vitamin Solution (1000x)

I am located in New Zealand, so shipping is pretty expensive. I want to avoid putting in a second order if I can.

Any help appreciated
Byron
Last edited by tokol0sh on Tue Feb 12, 2019 10:26 am, edited 1 time in total.

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TCGator

 
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Re: Getting Started

by TCGator » Tue Dec 18, 2018 8:04 pm

Thiamine and myo-inositol are the two most common vitamins in TC. I know that thiamine should be available at your local pharmacy/store. What about myo-inositol?

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tokol0sh

 
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Re: Getting Started

by tokol0sh » Fri Dec 21, 2018 11:05 am

Thanks for the reply. I added the vitamins to my order.

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tokol0sh

 
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Re: Getting Started

by tokol0sh » Wed Jan 23, 2019 1:06 pm

Just a bit of an update:
All my supplies are in and I have managed to repair an older autoclave from my fathers work (it had a loose connector and a blown fuse).

For my first batch I mixed up 175mL of 1/3 MS with vitamins (I guess the vitamins are at 1/3 now too?). This is enough to do 7 containers with 25mL each. I was going to add PPM but the volume needed was a bit small for me to measure accurately.

I dont have a laminar flow hood so just used an old wooden shipping crate I had around and lined it with plastic and sprayed it down with bleach about (with everything inside) about 30min before starting. Sterilized 14 flytrap seeds individually, I will definitely be using a coffee filter or similar next time...

The seeds were placed in the containers, 2 per container. Some seeds were place on the surface others submerged in the gel. Not sure if that matters?

Now to wait and see if there is any contamination and then if the seeds germinate.

Image


Image


Image

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tokol0sh

 
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Re: Getting Started

by tokol0sh » Mon Feb 11, 2019 8:21 am

About 2.5 weeks in, and no sign of contamination yet. No seeds have germinated yet. Got the seeds about 18" below a "300W" full spectrum LED grow light with a temperature of around 25-30C. Hopefully the seeds do alright...

I also mixed up a new batch of media:
1/3MS with full vitamins (I might have added too much of the MS vitamins solution so it might actually be 1.25
Vitamins)
1mL/L PPM

I made up a sterilization mix with 4% PPM and 1/3MS. This was used to try to sterilize some VFT cuttings, a sarracenia seedling and some drosera seeds. So far this is looking good. The VFT cuttings are still green (they have gone black where they were cut, but I suspect that was because they were cut with scissors and not a sharp scalpel), the sarracenia seedling has grown 2 new pitchers in about 2 weeks and the drosera seeds germinated in about 1 week. So far, I cant see any contamination.

This how that experiment is going:
Image



Here is my grow chamber:
Image

I will keep posting updates as things progress.

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jasonh


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