DNA Transfer to Cultured CellsKatya Ravid, R. Ian Freshney DNA transfer to cultured cells Edited by Katya Ravid and R. Ian Freshney Rapid advances in DNA transfer technology have transformed many disciplines, ranging from molecular genetics to biotechnology. Scientists now have the means to introduce copies of genes into different cell types, then detect the expression of these genes in the cell. It is now possible to regulate cell growth that may lead to cancer, develop new biopharmaceuticals, and apply knowledge about the role of genes in cell processes to basic research in molecular genetics. DNA Transfer to Cultured Cells is the first quick reference to all of the established techniques for the transfer of genetic material to cells in vitro. Featuring contributions by leading researchers in the field, this detailed guide walks the reader through a variety of DNA transfer methods, describes their application to specific cell types, and integrates aspects of molecular biology with tissue culture. Offering overviews and detailed protocols for the techniques under discussion in each of its sections, this book covers an exceptionally broad array of topics, including: * Viral infection * Electroporation * Phosphate precipitation * DEAE Dextran * Liposomes * Yeast artificial chromosomes (YACs) * Whole chromosome transfer * Enhanced expression. Special sections at the end of each chapter list suppliers for necessary reagents and materials. This easy-to-use, self-contained guide addresses key developments of recent years as well as emerging trends in DNA transfer. For practical applications in cell biology, genetics, heredity, biotechnology, or evolution, DNA Transfer to Cultured Cells is a unique and unparalleled resource. |
Contents
Preface | vii |
Baculovirus Expression and the Study of | 27 |
Electroporation of DNA into Cultured Cell Lines | 55 |
DNA Transfer to BlastulaDerived Cultures | 69 |
Transfection of Rat Adipose Cells | 93 |
Calcium Phosphate Transfection | 111 |
Optimization of Calcium Phosphate | 125 |
Strontium Phosphate Transfection | 147 |
Cationic Liposomes | 194 |
5 | 207 |
Transfection of Mammalian Cells with Yeast | 213 |
Mapping Human Senescence Genes Using | 237 |
Materials and Suppliers | 264 |
Methotrexate and DHFR Amplification | 265 |
List of Suppliers | 283 |
DEAEDextran Transfection | 179 |
Common terms and phrases
Acad Sci USA acid activity adipose cells agarose aliquots amplification antibody assay B-galactosidase baculovirus Biol buffer calcium phosphate cell culture cell lines centrifuge clones co-transfected CO₂ colonies concentration containing cultured cells DHFR dilution DMEM electroporation enzyme ethanol expression vector fibroblasts filter flask Freshney genetic genome Gibco BRL GLUT4 glycerol growth medium human chromosome hybrids immortal Incubate infection insulin isolated lipid Lipofectamine Lipofection liposomes mammalian cells marker Materials Aseptically prepared method methotrexate mg/ml microcell molecular monochromosome Natl Acad Sci Nonsterile PBSA plasmid plasmid DNA plate precipitate Proc Natl Acad producer cell protein Protocol Reagents and Materials recombinant replication reporter gene Resuspend retrovirus room temperature Section selection medium senescence Sigma solution staining sterile supernatant telomerase tion tissue culture titers transfected cells transfection transfection efficiency transgenic trypsin tubes viral virions virus YAC DNA yeast zebrafish µg/ml