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
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Page 15
... resuspend the cell pellet in 20 ml of 50 mM EDTA, pH 8.0, to wash the cells. Collect cells by spinning again for 5 min as before and decant supernatant. (4) Resuspend cells in 6 ml 50 mM EDTA, pH 8.0, and mix in 160 ml 10 mg/ml ...
... resuspend the cell pellet in 20 ml of 50 mM EDTA, pH 8.0, to wash the cells. Collect cells by spinning again for 5 min as before and decant supernatant. (4) Resuspend cells in 6 ml 50 mM EDTA, pH 8.0, and mix in 160 ml 10 mg/ml ...
Page 16
... resuspending in 2 ml of 200 mM NaCl, 10 mM Tris, pH 7.2, 100 mM EDTA. (5) Collect the cells by centrifuging at 4000g for 10 min. (6) Discard supernatant and resuspend cells in 0.5 ml of 200 mM NaCl, 10 mM Tris, pH 7.2, 100 mM EDTA by ...
... resuspending in 2 ml of 200 mM NaCl, 10 mM Tris, pH 7.2, 100 mM EDTA. (5) Collect the cells by centrifuging at 4000g for 10 min. (6) Discard supernatant and resuspend cells in 0.5 ml of 200 mM NaCl, 10 mM Tris, pH 7.2, 100 mM EDTA by ...
Page 34
... Resuspend the cells by pipetting in 2 ml of Sorbitol-EDTA. Collect the cells by a 5-min centrifugation at 3000g at 4°C. Discard the supernatant. (4) Resuspend the cells by pipetting in 1 ml of Sorbitol-EDTA-3mercaptoethanol, then ...
... Resuspend the cells by pipetting in 2 ml of Sorbitol-EDTA. Collect the cells by a 5-min centrifugation at 3000g at 4°C. Discard the supernatant. (4) Resuspend the cells by pipetting in 1 ml of Sorbitol-EDTA-3mercaptoethanol, then ...
Page 35
... resuspend the cells by pipetting in 50 ml of EDTA-3mercaptoethanol. Incubate for 20 min at room temperature with agitation (=100 rpm). Collect the cells by a 10-min centrifugation at 3000g at 4°C. Discard the supernatant in a fumehood ...
... resuspend the cells by pipetting in 50 ml of EDTA-3mercaptoethanol. Incubate for 20 min at room temperature with agitation (=100 rpm). Collect the cells by a 10-min centrifugation at 3000g at 4°C. Discard the supernatant in a fumehood ...
Page 36
... Resuspend the cells by pipetting in 20 ml of Sorbitol-EDTA. Collect the cells by a 10-min centrifugation at 3000g at 4°C. Discard the supernatant in a fumehood. (5) Resuspend the cells by pipetting in 20 ml of Novozym solution. Incubate ...
... Resuspend the cells by pipetting in 20 ml of Sorbitol-EDTA. Collect the cells by a 10-min centrifugation at 3000g at 4°C. Discard the supernatant in a fumehood. (5) Resuspend the cells by pipetting in 20 ml of Novozym solution. Incubate ...
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