Cell culture techniques allow a variety of molecular and cell biological questions to be addressed, offering physiological conditions whilst avoiding the use of laboratory animals. In addition to basic techniques, a wide range of specialised practical protocols covering the following areas are included: cell proliferation and death, in-vitro models for cell differentiation, in-vitro models for toxicology and pharmacology, industrial application of animal cell culture, genetic manipulation and analysis of human and animal cells in culture.

This manual describes the technique of differential-display reverse transcription-PCR. It contains step-by-step instructions, practical hints and experimental strategies for every protocol.

The present new version of this popular laboratory manual is at the same time the first one of this text in the English language - and this makes me even a little proud, as it reminds me of probably the first collection of monoclonal recipes in English, written by myself, which circulated for a couple of years in many laboratories. In the meantime many researchers have put enormous effort into improving methods for monoclonal antibody production. The proce dures have become more and more standardized and by this have more and more lost the character of magic secrets. Hinrich Peters and Horst...

UV-VIS spectroscopy is one of the oldest methods in molecular spectroscopy. The definitive formulation of the Bouguer-Lambert- Beer law in 1852 created the basis for the quantitative evaluation of absorption measurements at an early date. This led firstly to colorimetry, then to photometry and finally to spectrophotometry. This evolution ran parallel with the development of detectors for measuring light intensities, i.e. from the human eye via the photo- element and photocell, to the photomultiplier and from the photo- graphic plate to the present silicon-diode detector both of which allow...

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...

Tropical and subtropical countries have become well aware of the fact, that they must make better use of their fruits. In spite of the favourable climatic conditions for the production of varieties of delicious fruits in such countries, continuously high tempemtures shorten the shelf-life of most fruits and fruit products. A tropical climate provides ideal conditions for mpid growth of spoilage microorganisms and for chemical reactions. Most of such reactions in fruits and fruit products are deteriomtive in nature causing high respiration rates, texture softening and spoilage of fruit. This...

The intriguing complexity, precision, and regulation of the wide range of biologi cal processes is determined by intricate mechanisms of molecular recognition. Their nature is under intense scrutiny. In addition to the well-appreciated interac tion of proteins either with amino acid or nucleotide sequences, the investigation of their interplay with carbohydrate elements of cellular glycoconjugates current ly exerts increasing attraction. In the group of carbohydrate-binding proteins, lec tins are distinguished from antibodies or ligand-affecting enzymes, according to the most recent...

Capillary electrophoresis (CE) is a brand-new analytical me- thod with the capability of solving many analytical separa- tion problems very fast and economically. This method gives new information about the investigated substances which can- not easily be obtained by other means. CE has become an established method only recently, but will be implemented in almost every analytical laboratory in industry, service units and academia in the near future. The most important fields of CE application are pharmaceutical and biochemical research and quality control. The authors have exhaustive...

This book is addressed to both new and experienced workers interested in the properties of enzymes or using enzymes as gene markers in areas such as biochemi stry, clinical pathology, zoology, botany, systematics, ecology, population genetics or natural resources management. The first chapter introduces into the classification, structure and size of enzymes and isozymes. The second chapter provides protocols to extract (iso)enzymes from microbial, plant, animal and human tissue and cells-respectively. Methods to extract enzymes from cell-organelles of plant and animal tissues are also...

In this laboratory "cook-book," the authors provide a concise guide to PCR-based techniques to quantify nucleic acids in biological and clinical samples using exclusively nonradioactive detection methods, e.g. HPLC, biotin and digoxigenin based protocols. Each method presentation also includes sections on theory, reagents, standards, applicability, limitations, and trouble shooting. In addition to the protocols, the authors also provide the necessary information on: general aspects of nucleic acid quantitation; design of PCR standards; mRNA purification; cDNA synthesis; solution...