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This generation has had the opportunity to witness remarkable changes in nucleic acid (DNA/RNA) amplification technologies over the past 20 years. These technologies have made significant contributions to the advancement of molecular biology, to the understanding of the pathology and to the diagnosis of diseases. One of the most infamous examples is PCR. Since the invention of PCR by Kari Mullis in the late 1980s, PCR has become the dominant DNA amplification technology. Many variations of PCR were described including reverse transcription (RT)-PCR for the detection of RNA targets, long-distance PCR, panhandle PCR for cloning and ligation-dependent PCR for mutation detection. However, PCR technology remains in its evolutionary phase and new variations will be developed to meet increasingly demanding research and diagnostic needs.
Several extensions and refinements of the conventional PCR technology are described in the book entitled DNA Amplification: Current Technologies and Applications (Horizon Bioscience, Norfolk, UK, 2004), edited by Vadim V Demidov and Natalia E Broude, both from the Center for Advanced Biotechnology, Boston University, MA, USA. This book, in its unique format, reviews the latest developments in DNA amplification technologies. Also, its detailed layout of experimental protocols allows readers to implement these textbook methods in actual laboratory benchwork. Furthermore, each chapter presents a number of applications in length to improve the reader's understanding of each methodology.
The book has devoted a large section to the recent improvements of PCR technology, spanning from recombinant thermostable DNA polymerase to the applications of PCR in specific formats such as on-chip PCR, multi-plex PCR and real-time PCR. Pavlov and coworkers described a series of chimeric DNA polymerases that contain a DNA-binding domain from topoisomerase V and a catalytic domain from DNA polymerase. These modifications enable the enzymes to function at a higher temperature (100°C) and to tolerate a higher concentration of inhibitors (inter-calating dye and salt). The chapter on multiplex PCR by Broude and coworkers highlighted a method for amplifying multiple target sequences simultaneously using a specially designed adaptor that can be added to target DNA fragments by ligation. As a result, the amplification can be carried out using a generic primer that is complementary to...