Nonmammalian Genomic Analysis: A Practical Guide
Offering detailed protocols for those needing to construct a variety of maps and isolate genes, this unique book is intended to popularize the new techniques of genome analysis derived from the Human Genome Project. The power of these new methods is often most striking when applied to problems outside of human genetics, particularly the nonmammalian systems on which many researchers focus. Many of these organisms are economically important and biologically rich.
Nonmammalian Genomic Analysis: A Practical Guide covers the "how to" aspects of preparation, handling, cloning, and analysis of large DNA and the creation of chromosome and genome maps. This lab manual facilitates the transfer of these technologies to small "low tech" environments and allows them to be used by those with no background in genome mapping or large-fragment cloning. Like having a local expert, this collection provides procedures for anyone, anywhere, and allows the replication of others' success.
* Includes detailed and clearly-written step-by-step protocols
* Evinces expected results and offers trouble shooting advice
* Provides techniques appropriate for small laboratories as well as those with limited resources
* Covers a broad variety of cloning systems, including single copy vectors
* Discusses a diverse range of organisms, from prokaryotes to eukaryotes, from single-celled organisms to highly complex organisms
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Chapter 1 Introduction to PulsedField Gels and Preparation and Analysis of Large DNA
Chapter 2 Electrophoretic Karyotyping in Fungi
Chapter 3 Isolation and Analysis of HighMolecularWeight DNA from Plants
Chapter 4 Generating and Using DNA Markers in Plants
Chapter 5 Genome Mapping of Protozoan Parasites by Linking Clones
Chapter 6 Macrorestriction Mapping and Analysis of Bacterial Genomes
Chapter 7 Cosmid Cloning with Small Genomes
Chapter 8 Construction of PI Artificial Chromosome PAC Libraries from Lower Vertebrates
Chapter 9 The Selection of ChromosomeSpecific DNA Clones from African Trypanosome Genomic Libraries
Chapter 10 Analysis of the Dictyostelium discoideum Genome
Chapter 11 Integrated Genome Mapping by Hybridization Techniques
agarose gel aliquots ampliﬁcation bacterial bacteriophage bands blot buffer cells centrifuge chDNAs chro chromosomes coli colonies containing contigs cosmid Dictyostelium DNA fragments DNA molecules DNA sequence EDTA efﬁciency electrophoretic karyotype ethidium bromide ﬁeld ﬁlters ﬁnal Final concentration ﬁrst ﬂanking gel electrophoresis gene Genet genomic DNA hybridization identiﬁed Incubate insert isolation karyotype lane large DNA ligation linear linking clones macrorestriction mapping markers method molecular NaCl Natl Notl Novozym Nucleic Acids Res nucleotide oligonucleotide PAC cloning partial digestion PFGE physical mapping plasmid plates plugs polymerase polymorphisms prepare primers probe procedure protocol protoplasts pulse pulsed-ﬁeld gel RAPD reaction repeat restriction digestion restriction endonuclease restriction enzyme restriction fragments restriction mapping resuspend RFLP room temperature samples selected separation Sﬁl solution Southern blot speciﬁc spheroplasts sterile strains sufﬁcient switch techniques tion trypanosome tube vector vector DNA yeast