Ulrich Finckh – författare
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2 produkter
2 produkter
557 kr
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The polymerase chain reaction (PCR) - an in Vitro techniques for producing large amounts of a specific DNA fragment - has rapidly become established as one of the most important, impressive and fascinating methods of molecular biology as well as clinical diagnostics. In the seven years since'the technique was published, it has had a major impact on medical research. However, as there are still problems in instruments, standardized protocols for diagnostic applications and unsolved difficulties to avoid cross-contaminations on the one hand and on the other hand the even present question of how to interpret the biological value of a PCR result, most clinicians prefer to further wait until these topics are clarified. It is the aim of this book to give the reader lab-proven protocols from experienced scientists as well as a general introduction to alternative DNA-amplification procedures and their possible usage such as the NASBA or LCR. This book is divided into four major parts to provide a theoretical (first and second section) and a practical framework for a better understanding of the new technology. In the first part we provide an up-to-date summary of basic problems in this rapidly evolving field. We demonstrate, for example how to use fixed tissue materials and how to quantify PCR products as well as how to prepare nucleic acids in a safe, convenient and proper way, or even how to sequence directly PCR products for the analysis of the DNA structure.
1 075 kr
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In 1985, Kary Mullis was driving late at night through the flowering buckeye to the ancient California redwood forest, cogitating upon new ways to sequence DNA. Instead he came upon a way to double the number of specific DNA modules, and to repeat the process essentially indefinitely. 1 He thought of using two oligonucleotide sequences, oppositely oriented, and a DNA polymerase enzyme, to double the number of DNA targets. Each product would thene become the target for the next reaction, effectively yielding a product which doubled in quantity with each repeated cycle. Like the chain reaction leading to nuclear fission, with each cycle event each initial reactant (neutron or DNA molecule) yields two similar products, each of which can serve as the initial reactant. The invention of this exponentially increasing amplification system quickly became known as the polymerase chain reaction (PCR). The tremendous sensitivity of PCR ultimately resides in the necessity for each of two specific oligonucleotide annealing reactions to occur at the same time in the proper orientation. The DNA annealing reaction is a very specific reaction. Single genes have been detected by hybridization of a DNA probe to chromosome preparations together with sensitive fluorescence microscopy. This is the equivalent to detecting a gene present in a single copy per cell genome. It is the combination of two such specific annealing reactions which makes possible the amplification needed to detect a single molecule with a specific DNA sequence in over 100,000 cell genomes.