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 83
Page 2
... DNA in an agarose gel is placed in an electric field, the DNA molecules become elongated and oriented with the ... fragments to be resolved. The goal is to achieve the highest band resolution with minimum run time. Separation of the desired ...
... DNA in an agarose gel is placed in an electric field, the DNA molecules become elongated and oriented with the ... fragments to be resolved. The goal is to achieve the highest band resolution with minimum run time. Separation of the desired ...
Page 4
... DNA molecules migrate in an order that does not reflect increasing size. Some molecules will migrate faster than ... fragments on gels of several different switch times. The decreased resolution accompanying longer switch times and the ...
... DNA molecules migrate in an order that does not reflect increasing size. Some molecules will migrate faster than ... fragments on gels of several different switch times. The decreased resolution accompanying longer switch times and the ...
Page 5
... DNA. The sizes of the largest molecules that could be resolved in pulsed-field gels using a constant switch interval are shown for DNAs of three different size ranges. Reproducing ... DNA Fragments” Small. Introduction to Pulsed-Field Gels 5.
... DNA. The sizes of the largest molecules that could be resolved in pulsed-field gels using a constant switch interval are shown for DNAs of three different size ranges. Reproducing ... DNA Fragments” Small. Introduction to Pulsed-Field Gels 5.
Page 6
... DNA Fragments” Small DNA Medium DNA Big DNA Parameter (1–50 kb) (50–2000 kb) (>2000 kb) Voltage gradient (V/cm) 9 6 2% Reorientation angle 120° 120° 106° Temperature 14°C 14°C 14°C Buffer 0.5X TBE 0.5X TBE 1X TAE Agarose concentration 1 ...
... DNA Fragments” Small DNA Medium DNA Big DNA Parameter (1–50 kb) (50–2000 kb) (>2000 kb) Voltage gradient (V/cm) 9 6 2% Reorientation angle 120° 120° 106° Temperature 14°C 14°C 14°C Buffer 0.5X TBE 0.5X TBE 1X TAE Agarose concentration 1 ...
Page 7
... DNA (on the order of 20%), while others can cause a complete failure to resolve the fragments of interest. Therefore, understanding the relationship of these factors to the migration of the DNA allows the researcher to optimize the ...
... DNA (on the order of 20%), while others can cause a complete failure to resolve the fragments of interest. Therefore, understanding the relationship of these factors to the migration of the DNA allows the researcher to optimize the ...
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