Nonmammalian Genomic Analysis: A Practical GuideBruce Birren, Eric Lai 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.
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From inside the book
Results 1-5 of 78
Page 13
... restriction digestion. For preparation of intact chromosomes for separation by electrophoresis, conventional lowmelting agarose is effective. Ill. Preparation. of. Nonmammalian. Chromosomes. A. General Principles Traditional techniques for ...
... restriction digestion. For preparation of intact chromosomes for separation by electrophoresis, conventional lowmelting agarose is effective. Ill. Preparation. of. Nonmammalian. Chromosomes. A. General Principles Traditional techniques for ...
Page 14
... digested with restriction enzymes after preparation, the low-melting agarose must be free of compounds that inhibit enzyme activity. (4) Once cell walls have been digested, rapidly perform any necessary washes and embed the resulting ...
... digested with restriction enzymes after preparation, the low-melting agarose must be free of compounds that inhibit enzyme activity. (4) Once cell walls have been digested, rapidly perform any necessary washes and embed the resulting ...
Page 18
... restriction mapping or preparative uses for cloning, high-molecular-weight ... digestion with agarase before performing other enzymatic treatments. The ... restriction enzyme digestions where the resulting fragments are expected to be ...
... restriction mapping or preparative uses for cloning, high-molecular-weight ... digestion with agarase before performing other enzymatic treatments. The ... restriction enzyme digestions where the resulting fragments are expected to be ...
Page 19
... digestion can be controlled by limiting the amount of enzyme added, the amount of magnesium added, or the amount of ... Restriction Enzymes Choosing the appropriate restriction enzymes for cloning, fingerprinting, or long-range mapping ...
... digestion can be controlled by limiting the amount of enzyme added, the amount of magnesium added, or the amount of ... Restriction Enzymes Choosing the appropriate restriction enzymes for cloning, fingerprinting, or long-range mapping ...
Page 22
... restriction digestion. Most difficulties from gel conditions arise either from inattention to the set up and maintenance of the equipment or from failure to control each of the gel parameters. For example, if the gel box is not level or ...
... restriction digestion. Most difficulties from gel conditions arise either from inattention to the set up and maintenance of the equipment or from failure to control each of the gel parameters. For example, if the gel box is not level or ...
Contents
1 | |
25 | |
61 | |
Chapter 4 Generating and Using DNA Markers in Plants | 75 |
Chapter 5 Genome Mapping of Protozoan Parasites by Linking Clones | 135 |
Chapter 6 Macrorestriction Mapping and Analysis of Bacterial Genomes | 165 |
Chapter 7 Cosmid Cloning with Small Genomes | 197 |
Chapter 8 Construction of PI Artificial Chromosome PAC Libraries from Lower Vertebrates | 223 |
Chapter 9 The Selection of ChromosomeSpecific DNA Clones from African Trypanosome Genomic Libraries | 257 |
Chapter 10 Analysis of the Dictyostelium discoideum Genome | 293 |
Chapter 11 Integrated Genome Mapping by Hybridization Techniques | 319 |
Index | 347 |
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Common terms and phrases
Acad AFLP agarose gel aliquots amplification bacterial bacteriophage bands blot buffer cells centrifuge cerevisiae chDNAs chromosomes coli colonies containing contigs cosmid Dictyostelium DNA fragments DNA molecules DNA sequence EDTA electrophoretic karyotype ethidium bromide filters Final concentration gel electrophoresis gene Genet genomic DNA hybridization identified Incubate insert isolation karyotype lane large DNA ligation linear linking clones markers method mg/ml molecular NaCl Natl Novozym Nucleic Acids Res nucleotide oligonucleotide PAC cloning partial digestion PFGE physical mapping plasmid plates plugs polymerase polymorphisms prepare primers probe procedure proteinase K 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 switch techniques tion Tris-HCl trypanosome tube V/cm vector vector DNA yeast yeast artificial chromosomes