I have a story to
share. I was a research scholar in Biotechnology in IIT Bombay few years back. I was doing an
experiment on cell death by annexin V-Propidium Iodide (PI) assay. I was using a
kit supplied by a commercial manufacturer. Incidentally after a series of lab
mates used it and when my turn came the crucial buffer of the assay was finished
though the annexin V antibody and PI was very much there. My advisor suggested
me to not to depend on the manufacturer but to make my own buffer. I was
clueless because the supplier did not provide the buffer composition. I searched
in internet for a possible composition source. Ultimately a widely used cell
biology laboratory protocol book came to my rescue. I found that a simple salt
CaCl2 and a HEPES buffer were the key components. I made the buffer
according to the instruction laid in the book and voila! utilized that buffer
for very many reactions later on.
I learnt this
lesson to scour for the buffer composition because my buffer was finished and
the company did not provide the ‘secret’ composition. If all company start doing
this research will be hampered. Protocols and buffer compositions should be
clearly mentioned. In fact earlier version of Maniatis’s Molecular Cloning – a
bible for cloning had a nice preface urging the ‘Scientific Illiterates’ to know
of the reaction components scientists were using.
