I bought this book on the recommendation of several people, and basically this is by far the most well known, modern book on the topic. It covers the history, business, laboratory setup and design, media selection, sterile technique and even a short section on Genetic Engineering with Gene Gun and/or Agrobacteria.
Frankly, all of the above information can be had rather easily online and it isn’t in depth enough to be helpful if you are really looking to pursue this as a business, but the real value of the book comes in its kind of encyclopedic listing of media choices for plant species, this is very helpful and can be a guide to the budding Tissue Culture-ist as they venture out. I recommend this book for people, like me, who are starting out in Tissue Culture and who need a guide or Bible to help them illuminate some of the uncertainty as they get started. Well worth it.
At Dallas Maker Space, we worked to try to use Plant Tissue Culture, we used the MicroClone Starter Kit. The kit is probably a good introduction to plant tissue culture, but because we were new to this, we didn’t know that different types of plants require different media, Orchid is particularly picky and actually can poison itself with its own blood.
The MicroClone kit comes with three types of agar starters, plants themselves have hormones that tell them when to root, or branch, it was found that these hormones can be used by us to have plants root or branch on command -a very useful tool, when cloning plants. The kit comes with a BA hormone for multiplication/branching and also the NAA hormone for rooting/growth. We started with the Multiplication branching media. Following the instructions, we microwaved some Distilled water, poured the agar and Hormone into it as well as added sugar, a necessary ingredient. Then we used our autoclave to sterilize all of the containers, with the previously poured agar in them, I carefully closed the lids, and yet kept them slightly open so that they will not pop under the pressure, I also put my tweezers and blade into the autoclave as well.
Orchid
After 3 weeks of waiting, we found that the Orchid gives a black color in the multi medium, after asking some more experiences BioHackers about this they told us this is the Orchid blood and it will poison them eventually, Orchid media typically contains activated charcoal as well as banana, – who knew.
Fungis
The above picture is of a rose stem, now the stem is still green and alive, but the fungis is over taking the area around it, perhaps it was not perfectly sterilized before being put into the container.
The above picture is of two Rose stems, they are not contanminated with fungis, and will wait for a detailed analysis later.
Lessons Learned:
(1) We washed all of the various plants Orchid and Rose together, this was a mistake and could have led to cross contamination
(2) We needed to Autoclave the cutting board, which we did not do and was another source of contamination
(3) Need special Media for Orchids and need to research proper media for each plant type before moving forward.
Josh Melnick's Biology Blog 