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.
|
From inside the book
Results 1-5 of 30
Page 12
... NaCl 4 M ().625 ml 1()() m M EDTA 0.5 M, pH 8.0 l{).() Iml 0.2% Na deoxycholate 0.1 g 0.5% sarcosyl, Na salt, H20 0.25 g in 38.75 ml This solution may by prepared ahead of time. Add egg white lysozyme to 1 mg/ml final concentration ...
... NaCl 4 M ().625 ml 1()() m M EDTA 0.5 M, pH 8.0 l{).() Iml 0.2% Na deoxycholate 0.1 g 0.5% sarcosyl, Na salt, H20 0.25 g in 38.75 ml This solution may by prepared ahead of time. Add egg white lysozyme to 1 mg/ml final concentration ...
Page 13
... NaCl Add H2O to bring final volume to 1 liter. Sterilize by autoclaving. C. Choice of Agarose Most PFGs are cast at a 1% concentration using standard agarose sold for DNA electrophoresis. The agarose should be certified for use in ...
... NaCl Add H2O to bring final volume to 1 liter. Sterilize by autoclaving. C. Choice of Agarose Most PFGs are cast at a 1% concentration using standard agarose sold for DNA electrophoresis. The agarose should be certified for use in ...
Page 16
... 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 pipetting. Briefly warm to 37°C ...
... 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 pipetting. Briefly warm to 37°C ...
Page 18
... (e.g., 50 mM NaCl). In the third method, used to produce very large restriction fragments, enzymes and buffers diffuse into the agarose plugs which 18 Jennifer S. Lee et al. IV. Enzymatic Reactions Using DNA–Agarose Plugs.
... (e.g., 50 mM NaCl). In the third method, used to produce very large restriction fragments, enzymes and buffers diffuse into the agarose plugs which 18 Jennifer S. Lee et al. IV. Enzymatic Reactions Using DNA–Agarose Plugs.
Page 32
... NaCl 10% (w/v) SDS e. 2 M Sorbitol (pH 5.8) f 14.4 M 3-mercaptoethanol g. Proteinase K 2 | Final concentration To prepare 10 ml 20 mg/ml Proteinase K Dissolve 200 mg proteinase K in 10 ml 50 mM TrisHCl (pH 7.8) h. Sodium deoxycholate ...
... NaCl 10% (w/v) SDS e. 2 M Sorbitol (pH 5.8) f 14.4 M 3-mercaptoethanol g. Proteinase K 2 | Final concentration To prepare 10 ml 20 mg/ml Proteinase K Dissolve 200 mg proteinase K in 10 ml 50 mM TrisHCl (pH 7.8) h. Sodium deoxycholate ...
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 |
Other editions - View all
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