Re: [DIYbio] Plant Tissue Culture first attempt Dallas DIY Bio

Its a good idea to replate as soon as the "blood" pools for optimal culturing. It acts as a visual aid to maintain peak media efficiency. Media isnt necessarily for convenience's sake since every orchid species requires slightly more or less extra ingredients (coconut endosperm, banana powder, kinetin, etc.) and its cheaper to just buy the activated charcoal in bulk and have the media blank. Phytotechnologies Laboratories is your friend :)

Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC


On Nov 26, 2016, at 6:48 PM, Sebastian S Cocioba <scocioba@gmail.com> wrote:

For orchids best is internodal segments from floral spikes. Ideally from a freshly grown spike that has yet to flower. Leaf tissue is a pain to regenerate from. Aaron Hicks has an orchid seed repository unlike any other. Surface sterilizing seeds is also hit or miss. Best is to hand pollinate a phaleonopsis orchid and take the immature seed pod before it ripens, surface sterilize the pod, and sow the seeds that are intrinsically sterile since the seeds develop from the inside out. Prepare for long haul cultures when it comes to orchids. Set it and forget it. :)

Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC


On Nov 26, 2016, at 4:54 PM, Josh Melnick <j.r.melnick@gmail.com> wrote:

Thanks for your help, this was very helpful, I already ordered Tobacco seeds. I was looking at some Orchid Flask videos and yes indeed they have the black agar, from charcoal in it. 

On Friday, November 25, 2016 at 8:03:22 PM UTC-6, Sebastian wrote:
Welcome to the wonderful world of plant tissue culture!!!! ::confetti::

Orchids bleed and their blood is toxic to them. Use activated charcoal in orchid media.

Don't start with random plants. Start with tobacco seeds and surface sterilize them. Sprout them and use the resulting aseptic seeds as leaf tissue stock for subsequent practice cultures. 

10% household bleach, drop of dish soap, and pre-wash the plant tissue with soap and water manually to get macro-particles off. Soak in bleach for 10-30mins depending on surface area and convoluted tissue structures. Rinse 3x in sterile distilled water. Buy a heap of 6" tweezers and use one tweezer per culture vessel to not spread contam culture to culture. 

Orchid media works best with coconut water, a banana, and sucrose. Phytotechlab.com sells everything you need. Plants From Test Tubes 4th edition was my bible when I started. Keep the thread alive and I'll chime in. Best of luck!!! 

Sebastian S. Cocioba
CEO & Founder
New York Botanics, LLC


On Nov 25, 2016, at 3:06 AM, Josh Melnick <j.r.m...@gmail.com> wrote:

We are working on building a space here in Dallas, so far our first attempt at Tissue Culture has been rather poor. 
About 65% of the vials have mold or fungis, we have quite a good autoclave here, so I will have to work and be much more careful in the future about contamination when putting the plants in the tubes. 

I put orchid and rose in the tubes, using multi solution from MicroClone. What is interesting is this black coloration that is coming from the orchids, I suppose this is some product of the plant's metabolism as it is not coming into the other rose plants. I probably should start a blog. 

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