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 38
Page 3
... lane) and yeast chromosomes (right lane) were separated with switch intervals of 45, 75, or 105 sec. Each pulsedfield gel can separate a certain size range of DNA molecules, and lengthening the switch time increases this size range ...
... lane) and yeast chromosomes (right lane) were separated with switch intervals of 45, 75, or 105 sec. Each pulsedfield gel can separate a certain size range of DNA molecules, and lengthening the switch time increases this size range ...
Page 7
... lanes of the gel and over the duration of the gel run. PFGE buffers are usually either Tris Borate (TBE) or Tris Acetate (TAE) buffers. TBE is preferred for routine use, since it requires changing less frequently than TAE. For ...
... lanes of the gel and over the duration of the gel run. PFGE buffers are usually either Tris Borate (TBE) or Tris Acetate (TAE) buffers. TBE is preferred for routine use, since it requires changing less frequently than TAE. For ...
Page 16
... lanes containing 0.5 pig DNA per lane. (2) Arrest cell growth either by chilling cultures by swirling flasks in ice water or by adding chloramphenicol to a final concentration of 0.2 mg/ml and continuing incubation for 1 hr ...
... lanes containing 0.5 pig DNA per lane. (2) Arrest cell growth either by chilling cultures by swirling flasks in ice water or by adding chloramphenicol to a final concentration of 0.2 mg/ml and continuing incubation for 1 hr ...
Page 34
... lanes. This protocol can be scaled up or down as need be, although instead of scaling down, it is just as easy to form fewer plugs at the end. (1) Grow YNN295 in 10 ml of YPD in a 125-ml flask at 30°C with agitation (200–300 rpm) to ...
... lanes. This protocol can be scaled up or down as need be, although instead of scaling down, it is just as easy to form fewer plugs at the end. (1) Grow YNN295 in 10 ml of YPD in a 125-ml flask at 30°C with agitation (200–300 rpm) to ...
Page 35
... lanes. Using this protocol, O. ulmi s.l. chDNAs can be prepared after treatment with Novozym 234, or after treatment with several other lytic enzyme preparations (Fig. 2.1). (1) Grow O. ulmi s.l. in 50 ml medium in a 125-ml flask with ...
... lanes. Using this protocol, O. ulmi s.l. chDNAs can be prepared after treatment with Novozym 234, or after treatment with several other lytic enzyme preparations (Fig. 2.1). (1) Grow O. ulmi s.l. in 50 ml medium in a 125-ml flask with ...
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