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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Page 34
... room temperature 5 min. (5) Add 40 pil of Zymolyase 100T solution (or 50 34 Ken Dewar et al.
... room temperature 5 min. (5) Add 40 pil of Zymolyase 100T solution (or 50 34 Ken Dewar et al.
Page 35
... room temperature (hours to months) in digestion solution or plug storage buffer. 2. Neurospora crassa and Other Fungi This is an adaptation of the N. crassa procedure of Orbach et al. (1988), and is a representative protocol for ...
... room temperature (hours to months) in digestion solution or plug storage buffer. 2. Neurospora crassa and Other Fungi This is an adaptation of the N. crassa procedure of Orbach et al. (1988), and is a representative protocol for ...
Page 63
... room temperature overnight. (2) Transfer leaves in a petri dish with 20–40 ml protoplast isolation buffer and cut the leaves in 1- to 2-mm strips parallel to the veins with razor blades starting near the midvein while leaving the ...
... room temperature overnight. (2) Transfer leaves in a petri dish with 20–40 ml protoplast isolation buffer and cut the leaves in 1- to 2-mm strips parallel to the veins with razor blades starting near the midvein while leaving the ...
Page 64
... room temperature. Monitor digestion occasionally under an inverted microscope. For tomato leaves from greenhouse-grown plants, the digestion requires approximately 6 hrs. (5) After 6 hrs, most of the leaf should be digested away ...
... room temperature. Monitor digestion occasionally under an inverted microscope. For tomato leaves from greenhouse-grown plants, the digestion requires approximately 6 hrs. (5) After 6 hrs, most of the leaf should be digested away ...
Page 86
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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