What do you remember from the experience? Many people report an eerie feeling about their stays in hospitals.
But try to do your job without it, and you will see what a difference that simple little technique has made. If you type in 'pcr song,' you get a lovely little ditty courtesy of Bio-Rad, which will rattle around in your brain like an insane cat in your garage.
When I stumbled on PCR in the spring ofI was trying to increase the demand for oligonucleotides, which before automation my laboratory had made by hand. Our new machine from my friend Ron Cook at Biosearch across the San Francisco bay had threatened job stability in the laboratory by doing what had taken us about three weeks to do, in eight hours—and it did it every eight hours, no breaks.
The demand went up by about a million and I didn't have to fire any of my fellow lab workers at Cetus. I was driving up a long and winding road between Cloverdale and Booneville in Mendocino County, heading for my weekend cabin. My girlfriend was asleep and I was functionally sober or the road would have proven my undoing but it was late at night and I was feeling weird.
Strange things had happened to me on before. Furtive old men in…what was that? I didn't see anything. Oligonucleotides are amazing little things, but using only one, it is not possible to physically locate a particular spot on human DNA.
If the human genome were random, a nucleotide oligomer would uniquely specify a position along the 6 to 7 billion bases in denatured human DNA. But it's not random, and any mer that is in there, is probably in there more than once, or at least some slightly different version is in there.
There was no way to know that for sure in the early eighties, and there are more complicated arguments for why this is so, but if you looked at gels of whole human DNA broken into restriction fragments and probed with mers, you saw a lot of smears.
No really sharp bands like the restriction digests of bacteriophage DNA that you could use as markers. So if you wanted to examine a human DNA sequence closely, you had to clone it. Chop up the DNA into pieces of several thousand base pairs, isolate each of those by growing them in a particular bacterial colony, figure out which colony contained your favorite piece, pick it off a plate and grow it up.
That was the magic of cloning, and it was magic. We all knew it. Even the janitors pushing the brooms through the laboratories at night could feel it.
In the late seventies, just as I started working for Cetus, a number of prominent molecular biologists convinced the rest of the field to hold off a little to ponder the safety issues. Conferences were called, laws were even passed in Cambridge, Massachusetts and Berkeley, California.
We were safely in Emeryville, where there were gambling houses, but few laws. No one could be sure that putting human genes into micro-organisms that could possibly infect humans, was such a good idea.
They never did figure it out, but by way of compromising, some strains of E.
Unconsciously combining the two problems, I started devising methods whereby oligonucleotides could be used to determine single base pair mutations from whole human DNA.
Pregnant mothers should not have to wait for the cloners, and the result of running gels and using radioactive probes on genomic DNA were fuzzy for reasons mentioned above. Fuzzy is not a comfortable basis for making a life or death decision.
Somebody needed to come up with a way to concentrate a single DNA locus in the presence of millions of similar but different DNA loci without the inevitable delay of cloning. It was going to happen tonight. That somebody was going to be me.Sister Papers Works from the Louvre Abu Dhabi: Dr Lamees Hamdan on the Bactrian Princess Cellulose.
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James Mack is an AssociateProfessor of Chemistry. After completing his Bachlor’s degree at Middlebury College in he went to graduate school at the University of New Hampshire where he conducted his doctoral research under the supervision of Glen P. Miller working in the area of fullerenes.
Distilled water tends to be acidic and can only be recommended as a way of drawing poisons out of the body. Once this is accomplished, the continued drinking of distilled water is a bad idea.
Water filtered through reverse osmosis tends to be neutral and is acceptable . Dr. Joseph M. Genco, Dr. David Neivandt and Dr. Haixuan Zou for accepting to spend their time and energy for the benefit of my thesis.
I would like to thank the Turkish Paper .