john_ny

In the Cold Hardy Bananas section, I saw a thread about, " Musa basjoo, then and now", and I thought of my experiences, then and now, with TC. When I read the thread, I saw that the time between the then and now, with the basjoo, was several months. The difference between my first contact with tissue culture (then), and now, is almost 30 years.
In the 1979 issue of, "Compact Fruit Tree", the journal of the International Dwarf Fruit Tree Association, is an article about tissue culture which includes the basic procedure, and some recipes for media. In those days, for the most part, you couldn't buy media; you had to obtain all of the chemical components, and mix them yourself. I expressed my interest to my wife, (an operating room supervisor) and she spoke to her boss (chief of surgery), and he said that he would get the chemicals for me from the lab. Then I had to measure the chemicals out, and some of the quantities are tiny, minute. (We're talking micro Moles & nano Moles, here) We needed a super accurate scale from a scientific supply house. They wouldn't sell it to individuals, (too often used for drugs) so my wife got the hospital to buy it, and she reimbursed them. Then, I had to have some containers to put the media in. I got baby food jars, from a neighbor, who had a small child. A pressure cooker was used to sterilize the jars and media. I didn't have a hood, for doing the transfers, so I had quite a bit of contamination, but I had one pear tree that made it, and is still in the yard today. There were lots of errors made then, by me, and the people who wrote the papers. (Some months later, I read where someone pointed out that some of the media components are heat-labile, and break down, or are destroyed, by steam sterilization.) Surprisingly though, things worked, anyway.
In subsequent tries, I either got the hospital lab to weigh out the media, or bought it pre-measured, and used my wife's OR autoclave for sterilizing, and the lab's laminar-flow hood for transfers.

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John