[理学]PCR2007.pdf

Polymerase Chain Reaction (PCR) ustcwhm@ustc.edu.cn Kary B. Mullis *One Friday night I was driving, ……. My girlfriend, Jennifer Barnett, was asleep. I was thinking. ……. *However, shocking to me, not one of my friends or colleagues would get excited over the potential for such a process. *In September I did my first experiment. I like to try the easiest possibilities first. So one night I put human DNA and the nerve growth factor primers in a little screw-cap tube with an O-ring and a purple top. I boiled for a few minutes, cooled, added about 10 units of DNA polymerase, closed the tube and left it at 37 °. It was exactly midnight on the ninth of September. *The first successful experiment happened on December 16th. I remember the date. It was the birthday of Cynthia, my former wife from Kansas City, who had encouraged me to write fiction and bore us two fine sons. I had strayed from Cynthia eventually to spend two tumultuous years with Jennifer. The Nobel Prize in Chemistry 1993 for contributions to the developments of methods within DNA-based chemistry for his invention of the polymerase chain reaction (PCR) method Kary B. Mullis La Jolla, CA, USA B:1944 PCR反应混合物 Mg2+ 和 Taq DNA 多聚酶 或 2+ Mn 和 rTth DNA 多聚酶 dCTP AmpErase dGTP （UNG酶） dUTP 引 物 dATP 1.Choice of Polymerases for PCR One of the important advances which allowed development of PCR was the availability of thermostable polymerases. This allowed initially added enzyme to survive temperature cycles approaching 100 °C. Thermostable DNA polymerases and their sources