Guy A. Caldwell, Shelli N. Williams, Kim A. Caldwell

Integrated Genomics

A Discovery-Based Laboratory Course

• Broschiertes Buch

Produktbeschreibung

Diese spannende, gut nachvollziehbare Einführung in das moderne Gebiet der Genomforschung ist eine gelungene Mischung ais methogologischen Informationen zur Molekularbiologie und Arbeitsanleitungen zur sinnvollen Benutzung online verfügbarer Datenbanken.

Produktdetails

• Verlag: Wiley & Sons
• 1. Auflage
• Seitenzahl: 248
• Erscheinungstermin: 1. September 2006
• Englisch
• Abmessung: 280mm x 210mm x 14mm
• Gewicht: 719g
• ISBN-13: 9780470095027
• ISBN-10: 0470095024
• Artikelnr.: 21539784

Autorenporträt

Guy A. Caldwell, Ph.D., is an Assistant Professor in the Department of Biological Sciences at The University of Alabama in Tuscaloosa, where since 1999, he has held an undergraduate professorial appointment from the Howard Hughes Medical Institute. In 2001, Dr. Caldwell was named a Basil O'Connor Scholar of The March of Dimes Birth Defects Foundation for his research into the molecular basis of childhood birth defects of the brain. Dr. Caldwell is a recipient of grants from The March of Dimes, National Institutes of Health, the Dystonia Medical Research Foundation, American Parkinson's Disease Association, Parkinson's Disease Foundation, and the National Parkinson Foundation. In January 2003, The Caldwell lab was selected as one of only 11 worldwide to represent the research goals of The Michael J. Fox Foundation for Parkinson's Research in their Protein Degradation Grant Initiative. For his combined teaching and research efforts, Dr. Caldwell was also chosen as the recipient of a 2003 CAREER award from the National Science Foundation, the most prestigious honor for young faculty bestowed by that organization. He is the author of 2 editions of a widely adopted textbook in biotechnology sold worldwide in 3 languages by Harcourt. He currently teaches courses in Molecular Genomics, Neuronal Signaling Mechanisms, General Biology, and an acclaimed seminar on the societal impact of the Human Genome Project. Shelli N. Williams, B.Sc., is a doctoral candidate in the Department of Biological Sciences at The University of Alabama in Tuscaloosa, where she has attended the university as an undergraduate and graduate student since 1997. Following her early graduation magna cum laude from the university, Ms. Williams began her graduate work in the laboratory of Dr. Guy A. Caldwell. She has experience teaching introductory biology courses to both major and non-major students and has served as teaching assistant for a senior level discovery-based genomics course funded by the Howard Hughes Medical Institute. Kim A. Caldwell, Ph.D. is an Adjunct Assistant Professor in the Department of Biological Sciences at The University of Alabama Dr. Caldwell serves as an administrative liaison for a 1.8 million dollar grant from the Howard Hughes Medical Institute to the Department of Biological Sciences at Alabama and is Director of the HHMI Rural Science Scholars program at Alabama. She has designed and taught courses in General Biology, a seminar on the societal impact of the Human Genome Project, and course is a entitled "The Language of Research" which she teaches jointly for Howard Hughes Research Interns at both Stillman College and The University of Alabama.

Inhaltsangabe

Preface. Author biographies. Acknowledgments. List of figures. 1
Introduction to basic laboratory genetics. 1.1 Transferring and handling C.
elegans. 1.2 Introduction to laboratory genetics. 2 Gene expression
analysis using transgenic animals. 2.1 Transgenic gene expression analysis
in C. elegans: lacZ staining. 2.2 Transgenic gene expression analysis in C.
elegans: GFP analysis. 3 Creation and testing of transgenic yeast for use
in protein-protein interaction screening. 3.1 Small-scale transformation of
S. cerevisiae. 3.2 Transformation of S. cerevisiae to test for non-specific
interaction. 3.3 Assaying for protein-protein interaction by reporter gene
expression. 4 Yeast two-hybrid screening. 4.1 Protein-protein interaction
screening of a C. elegans cDNA library. 4.2 Assaying for protein-protein
interaction by reporter gene expression. 5 Isolation and identification of
interacting proteins. 5.1 Preparation of electrocompetent E. coli. 5.2
Isolation of DNA from yeast and electroporation of E. coli. 5.3 Small-scale
isolation of plasmid DNA from E. coli: the mini-prep. 5.4 Sequencing of
two-hybrid library plasmid DNA vectors. 6 Using bioinformatics in modern
science. 6.1 DNA sequence chromatogram. 6.2 BLASTing your sequence. 6.3
Evaluating sequence results and choosing an RNAi target. 6.4 Bioinformatics
practice questions. 7 Generation of an RNAi vector. 7.1 Small-scale
isolation of genomic DNA from C. elegans. 7.2 PCR amplification of target
gene sequence from C. elegans genomic DNA. 7.3 Preparations for cloning to
generate RNAi vector. 7.3.1 Agarose gel electrophoresis. 7.3.2 Removal of
dNTPs from PCR reaction. 7.3.3 Restriction enzyme digestion of PCR product
and C. elegans RNAi vector. 7.4 Gel purification of DNA and ligation of
vector and PCR-amplified DNA. 7.4.1 Preparative agarose gel
electrophoresis. 7.4.2 Gel purification of DNA from agarose gel. 7.4.3
Ligation of vector and PCR-amplified DNA. 7.5 Transformation of ligation
reactions. 7.6 PCR screening of transformation colonies. 7.7 Small-scale
isolation of plasmid DNA from E. coli: the mini-prep. 7.8 Verifying
successful ligation by restriction digestion. 8 RNA-mediated interference
by bacterial feeding. 8.1 Preparation of RNAi-feeding bacteria for
transformation. 8.2 Media preparation for RNAi feeding. 8.3 Transformation
of RNAi-feeding strain HT115(DE3). 8.4 RNA interference by bacterial
feeding of C. elegans. 8.5 Analyzing effects of dsRNAi. 8.5.1 Assaying for
sterility (Ste) or embryonic lethality (Emb). 8.5.2 Assaying for growth
effect. 8.5.3 Assaying for morphological effects. 8.5.4 Assaying for
general neuromuscular effects. 8.5.5 Assaying for specific neuronal
effects. 8.5.6 Assaying for dauer formation. Appendix I Recombinational
cloning. AI.1 Isolation of genomic DNA from C. elegans. AI.2 PCR
amplification of target gene sequence from C. elegans genomic DNA. AI.3
Agarose gel electrophoresis and clean-up of PCR reaction. AI.4 Entry vector
cloning. AI.5 Small-scale isolation of plasmid DNA from E. coli: the
mini-prep. AI.6 Destination vector cloning. AI.7 Small-scale isolation of
plasmid DNA from E. coli: the mini-prep. Appendix II Recipes and media
preparation. Solution recipes. Media preparation. Appendix III Sterile
techniques and worm protocols. Sterile techniques. Worm protocols. Appendix
IV Mutant C. elegans phenotypes. Appendix V Vector maps. Subject index.