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2.
Nature ; 535(7612): 435-439, 2016 07 21.
Artículo en Inglés | MEDLINE | ID: mdl-27409808

RESUMEN

Recent work has underscored the importance of the microbiome in human health, and has largely attributed differences in phenotype to differences in the species present among individuals. However, mobile genes can confer profoundly different phenotypes on different strains of the same species. Little is known about the function and distribution of mobile genes in the human microbiome, and in particular whether the gene pool is globally homogenous or constrained by human population structure. Here, we investigate this question by comparing the mobile genes found in the microbiomes of 81 metropolitan North Americans with those of 172 agrarian Fiji islanders using a combination of single-cell genomics and metagenomics. We find large differences in mobile gene content between the Fijian and North American microbiomes, with functional variation that mirrors known dietary differences such as the excess of plant-based starch degradation genes found in Fijian individuals. Notably, we also observed differences between the mobile gene pools of neighbouring Fijian villages, even though microbiome composition across villages is similar. Finally, we observe high rates of recombination leading to individual-specific mobile elements, suggesting that the abundance of some genes may reflect environmental selection rather than dispersal limitation. Together, these data support the hypothesis that human activities and behaviours provide selective pressures that shape mobile gene pools, and that acquisition of mobile genes is important for colonizing specific human populations.


Asunto(s)
Transferencia de Gen Horizontal/genética , Interacción Gen-Ambiente , Variación Genética/genética , Metagenómica , Microbiota/genética , Selección Genética/genética , Bacteriófagos/genética , Estudios de Cohortes , Elementos Transponibles de ADN/genética , Dieta , Fiji , Pool de Genes , Humanos , América del Norte , Plásmidos/genética , Recombinación Genética/genética , Análisis de la Célula Individual
3.
Infect Genet Evol ; 11(8): 2011-9, 2011 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21964598

RESUMEN

Dengue virus currently causes 50-100 million infections annually. Comprehensive knowledge about the evolution of Dengue in response to selection pressure is currently unavailable, but would greatly enhance vaccine design efforts. In the current study, we sequenced 187 new dengue virus serotype 3 (DENV-3) genotype III whole genomes isolated from Asia and the Americas. We analyzed them together with previously-sequenced isolates to gain a more detailed understanding of the evolutionary adaptations existing in this prevalent American serotype. In order to analyze the phylogenetic dynamics of DENV-3 during outbreak periods; we incorporated datasets of 48 and 11 sequences spanning two major outbreaks in Venezuela during 2001 and 2007-2008, respectively. Our phylogenetic analysis of newly sequenced viruses shows that subsets of genomes cluster primarily by geographic location, and secondarily by time of virus isolation. DENV-3 genotype III sequences from Asia are significantly divergent from those from the Americas due to their geographical separation and subsequent speciation. We measured amino acid variation for the E protein by calculating the Shannon entropy at each position between Asian and American genomes. We found a cluster of seven amino acid substitutions having high variability within E protein domain III, which has previously been implicated in serotype-specific neutralization escape mutants. No novel mutations were found in the E protein of sequences isolated during either Venezuelan outbreak. Shannon entropy analysis of the NS5 polymerase mature protein revealed that a G374E mutation, in a region that contributes to interferon resistance in other flaviviruses by interfering with JAK-STAT signaling was present in both the Asian and American sequences from the 2007-2008 Venezuelan outbreak, but was absent in the sequences from the 2001 Venezuelan outbreak. In addition to E, several NS5 amino acid changes were unique to the 2007-2008 epidemic in Venezuela and may give additional insight into the adaptive response of DENV-3 at the population level.


Asunto(s)
Virus del Dengue/clasificación , Virus del Dengue/genética , Dengue/epidemiología , Dengue/virología , Genoma Viral , Mutación , Américas/epidemiología , Sustitución de Aminoácidos , Animales , Secuencia de Bases , Teorema de Bayes , Dengue/genética , Evolución Molecular , Genotipo , Humanos , Datos de Secuencia Molecular , Filogenia , Serotipificación , Venezuela/epidemiología
4.
mBio ; 2(1): e00342-10, 2011.
Artículo en Inglés | MEDLINE | ID: mdl-21304167

RESUMEN

Cryptococcus gattii recently emerged as the causative agent of cryptococcosis in healthy individuals in western North America, despite previous characterization of the fungus as a pathogen in tropical or subtropical regions. As a foundation to study the genetics of virulence in this pathogen, we sequenced the genomes of a strain (WM276) representing the predominant global molecular type (VGI) and a clinical strain (R265) of the major genotype (VGIIa) causing disease in North America. We compared these C. gattii genomes with each other and with the genomes of representative strains of the two varieties of Cryptococcus neoformans that generally cause disease in immunocompromised people. Our comparisons included chromosome alignments, analysis of gene content and gene family evolution, and comparative genome hybridization (CGH). These studies revealed that the genomes of the two representative C. gattii strains (genotypes VGI and VGIIa) are colinear for the majority of chromosomes, with some minor rearrangements. However, multiortholog phylogenetic analysis and an evaluation of gene/sequence conservation support the existence of speciation within the C. gattii complex. More extensive chromosome rearrangements were observed upon comparison of the C. gattii and the C. neoformans genomes. Finally, CGH revealed considerable variation in clinical and environmental isolates as well as changes in chromosome copy numbers in C. gattii isolates displaying fluconazole heteroresistance.


Asunto(s)
Criptococosis/inmunología , Criptococosis/microbiología , Cryptococcus gattii/genética , Variación Genética , Genoma Bacteriano , Animales , Antifúngicos/farmacología , Cryptococcus gattii/clasificación , Cryptococcus gattii/efectos de los fármacos , Cryptococcus gattii/aislamiento & purificación , Brotes de Enfermedades , Evolución Molecular , Femenino , Genotipo , Interacciones Huésped-Patógeno , Humanos , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , América del Norte/epidemiología , Filogenia
5.
Genome Res ; 9(10): 994-1001, 1999 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-10523528

RESUMEN

Human genome sequencing is accelerating rapidly. Multiple genome maps link this sequence to problems in biology and clinical medicine. Because each map represents a different aspect of the structure, content, and behavior of human chromosomes, these fundamental properties must be integrated with the genome to understand disease genes, cancer instability, and human evolution. Cytogenetic maps use 400-850 visible band landmarks and are the primary means for defining prenatal defects and novel cancer breakpoints, thereby providing simultaneous examination of the entire genome. Recent genetic, physical, and transcript maps use PCR-based landmarks called sequence-tagged sites (STSs). We have integrated these genome maps by anchoring the human cytogenetic to the STS-based genetic and physical maps with 1021 STS-BAC pairs at an average spacing of approximately 1 per 3 Mb. These integration points are represented by 872 unique STSs, including 642 polymorphic markers and 957 bacterial artificial chromosomes (BACs), each of which was localized on high resolution fluorescent banded chromosomes. These BACs constitute a resource that bridges map levels and provides the tools to seamlessly translate questions raised by genomic change seen at the chromosomal level into answers based at the molecular level. We show how the BACs provide molecular links for understanding human genomic duplications, meiosis, and evolution, as well as reagents for conducting genome-wide prenatal diagnosis at the molecular level and for detecting gene candidates associated with novel cancer breakpoints.


Asunto(s)
Cromosomas Bacterianos , Genoma Humano , Mapeo Cromosómico , Cromosomas Humanos Par 11/ultraestructura , Humanos , Hibridación Fluorescente in Situ , Modelos Genéticos , Mapeo Físico de Cromosoma , Reproducibilidad de los Resultados , Lugares Marcados de Secuencia
6.
Nat Genet ; 23(1): 104-7, 1999 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-10471509

RESUMEN

Early outgrowth of the vertebrate embryonic limb requires signalling by the apical ectodermal ridge (AER) to the progress zone (PZ), which in response proliferates and lays down the pattern of the presumptive limb in a proximal to distal progression. Signals from the PZ maintain the AER until the anlagen for the distal phalanges have been formed. The semidominant mouse mutant dactylaplasia (Dac) disrupts the maintenance of the AER, leading to truncation of distal structures of the developing footplate, or autopod. Adult Dac homozygotes thus lack hands and feet except for malformed single digits, whereas heterozygotes lack phalanges of the three middle digits. Dac resembles the human autosomal dominant split hand/foot malformation (SHFM) diseases. One of these, SHFM3, maps to chromosome 10q24 (Refs 6,7), which is syntenic to the Dac region on chromosome 19, and may disrupt the orthologue of Dac. We report here the positional cloning of Dac and show that it belongs to the F-box/WD40 gene family, which encodes adapters that target specific proteins for destruction by presenting them to the ubiquitination machinery. In conjuction with recent biochemical studies, this report demonstrates the importance of this gene family in vertebrate embryonic development.


Asunto(s)
Extremidades/embriología , Deformidades Congénitas de las Extremidades/genética , Proteínas/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Mapeo Cromosómico , Proteínas F-Box , Heterocigoto , Humanos , Mesodermo/metabolismo , Ratones , Ratones Endogámicos BALB C , Modelos Genéticos , Datos de Secuencia Molecular , Familia de Multigenes , Mutación , Homología de Secuencia de Aminoácido , Factores de Tiempo , Distribución Tisular
7.
Nat Genet ; 22(4): 388-93, 1999 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-10431246

RESUMEN

A physical map of the mouse genome is an essential tool for both positional cloning and genomic sequencing in this key model system for biomedical research. Indeed, the construction of a mouse physical map with markers spaced at an average interval of 300 kb is one of the stated goals of the Human Genome Project. Here we report the results of a project at the Whitehead Institute/MIT Center for Genome Research to construct such a physical map of the mouse. We built the map by screening sequenced-tagged sites (STSs) against a large-insert yeast artificial chromosome (YAC) library and then integrating the STS-content information with a dense genetic map. The integrated map shows the location of 9,787 loci, providing landmarks with an average spacing of approximately 300 kb and affording YAC coverage of approximately 92% of the mouse genome. We also report the results of a project at the MRC UK Mouse Genome Centre targeted at chromosome X. The project produced a YAC-based map containing 619 loci (with 121 loci in common with the Whitehead map and 498 additional loci), providing especially dense coverage of this sex chromosome. The YAC-based physical map directly facilitates positional cloning of mouse mutations by providing ready access to most of the genome. More generally, use of this map in addition to a newly constructed radiation hybrid (RH) map provides a comprehensive framework for mouse genomic studies.


Asunto(s)
Cromosomas Artificiales de Levadura , Genoma , Ratones/genética , Mapeo Físico de Cromosoma , Animales , Mapeo Cromosómico , Mapeo Contig , Marcadores Genéticos , Modelos Genéticos
8.
Genome Res ; 9(5): 514-23, 1999 May.
Artículo en Inglés | MEDLINE | ID: mdl-10330132

RESUMEN

We have established a collection of strong molecular cytogenetic markers that span the mouse autosomes and X chromosome at an average spacing of one per 19 Mb and identify 127 distinct band landmarks. In addition, this Mouse Molecular Cytogenetic Resource relates the ends of the genetic maps to their chromosomal locations. The resource consists of 157 bacterial artificial chromosome (BAC) clones, each of which identifies specific mouse chromosome bands or band borders, and 42 of which are linked to genetic markers that define the centromeric and telomeric ends of the Whitehead/MIT recombinational maps. In addition, 108 randomly selected and 6 STS-linked BACs have been assigned to single chromosome bands. We have also developed a high-resolution fluorescent reverse-banding technique for mouse chromosomes that allows simultaneous localization of probes by fluorescence in situ hybridization (FISH) with respect to the cytogenetic landmarks. This approach integrates studies of the entire mouse genome. Moreover, these reagents will simplify gene mapping and analyses of genomic fragments in fetal and adult mouse models. As shown with the MMU16 telomeric marker for the trisomy 16 mouse model of Down syndrome, these clones can obviate the need for metaphase analyses. The potential contribution of this resource and associated methods extends well beyond mapping and includes clues to understanding mouse chromosomes and their rearrangements in cancers and evolution. Finally it will facilitate the development of an integrated view of the mouse genome by providing anchor points from the genetic to the cytogenetic and functional maps of the mouse as we attempt to understand mutations, their biological consequences, and gene function.


Asunto(s)
Mapeo Cromosómico/métodos , Cromosomas Bacterianos/genética , Marcadores Genéticos/genética , Animales , Bandeo Cromosómico/métodos , Hibridación Fluorescente in Situ/métodos , Cariotipificación , Ratones , Ratones Endogámicos , Telómero/genética
9.
Nat Genet ; 19(3): 274-8, 1998 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-9662403

RESUMEN

Pudgy (pu) homozygous mice exhibit clear patterning defects at the earliest stages of somitogenesis, resulting in adult mice with severe vertebral and rib deformities. By positional cloning and complementation, we have determined that the pu phenotype is caused by a mutation in the delta-like 3 gene (Dll3), which is homologous to the Notch-ligand Delta in Drosophila. Histological and molecular marker analyses show that the pu mutation disrupts the proper formation of morphological borders in early somite formation and of rostral-caudal compartment boundaries within somites. Viability analysis also indicates an important role in early development. The results point to a key role for a Notch-signalling pathway in the initiation of patterning of vertebrate paraxial mesoderm.


Asunto(s)
Glicosiltransferasas , Proteínas de la Membrana/genética , Proteínas de la Membrana/fisiología , Mutación , Somitos/fisiología , Empalme Alternativo , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Mapeo Cromosómico , ADN Complementario , Péptidos y Proteínas de Señalización Intracelular , Ratones , Datos de Secuencia Molecular , Proteínas/metabolismo
10.
Nature ; 389(6652): 722-5, 1997 Oct 16.
Artículo en Inglés | MEDLINE | ID: mdl-9338782

RESUMEN

The mouse syndactylism (sm) mutation impairs some of the earliest aspects of limb development and leads to subsequent abnormalities in digit formation. In sm homozygotes, the apical ectodermal ridge (AER) is hyperplastic by embryonic day 10.5, leading to abnormal dorsoventral thickening of the limb bud, subsequent merging of the skeletal condensations that give rise to cartilage and bone in the digits, and eventual fusion of digits. The AER hyperplasia and its effect on early digital patterning distinguish sm from many other syndactylies that result from later failure of cell death in the interdigital areas. Here we use positional cloning to show that the gene mutated in sm mice encodes the putative Notch ligand Serrate. The results provide direct evidence that a Notch signalling pathway is involved in the earliest stages of limb-bud patterning and support the idea that an ancient genetic mechanism underlies both AER formation in vertebrates and wing-margin formation in flies. In addition to cloning the sm gene, we have mapped three modifiers of sm, for which we suggest possible candidate genes.


Asunto(s)
Proteínas Portadoras/genética , Proteínas de la Membrana/genética , Sindactilia/genética , Secuencia de Aminoácidos , Animales , Mapeo Cromosómico , Cruzamientos Genéticos , Ectodermo/metabolismo , Exones , Extremidades/embriología , Femenino , Expresión Génica , Ligamiento Genético , Glicina/química , Péptidos y Proteínas de Señalización Intracelular , Intrones , Masculino , Ratones , Ratones Endogámicos BALB C , Ratones Mutantes , Datos de Secuencia Molecular , Fenotipo , Homología de Secuencia de Aminoácido , Sindactilia/embriología
11.
Proc Natl Acad Sci U S A ; 93(13): 6297-301, 1996 Jun 25.
Artículo en Inglés | MEDLINE | ID: mdl-8692809

RESUMEN

We have constructed a physical map of human chromosome 22q using bacterial artificial chromosome (BAC) clones. The map consists of 613 chromosome 22-specific BAC clones that have been localized and assembled into contigs using 452 landmarks, 346 of which were previously ordered and mapped to specific regions of the q arm of the chromosome by means of chromosome 22-specific yeast artificial chromosome clones. The BAC-based map provides immediate access to clones that are stable and convenient for direct genome analysis. The approach to rapidly developing marker-specific BAC contigs is relatively straightforward and can be extended to generate scaffold BAC contig maps of the rest of the chromosomes. These contigs will provide substrates for sequencing the entire human genome. We discuss how to efficiently close contig gaps using the end sequences of BAC clone inserts.


Asunto(s)
Cromosomas Humanos Par 22 , Línea Celular , Mapeo Cromosómico , Cromosomas Bacterianos , Biblioteca Genómica , Humanos
12.
Genomics ; 34(2): 213-8, 1996 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-8661051

RESUMEN

We have constructed an arrayed human genomic BAC library with approximately 4x coverage that is represented by 96,000 BAC clones with average insert size of nearly 140 kb. A new BAC vector that allows color-based positive screening to identify transformants with inserts has increased BAC cloning efficiency. The library was gridded onto hybridization filters at high density for efficient identification of BAC clones by colony hybridization. The library was also formulated into characteristic DNA pools to allow for PCR screening of the library for STS content. We have characterized the library mainly by screening with more than 300 different landmarks that include cDNA, STSs, and cosmid clones. We describe methods for using BAC clones and discuss the implications for genome characterization, mapping, and sequencing.


Asunto(s)
Cromosomas Bacterianos , Biblioteca Genómica , Línea Celular , Clonación Molecular/métodos , Cósmidos , ADN/análisis , Marcadores Genéticos , Técnicas Genéticas , Humanos , Masculino , Reacción en Cadena de la Polimerasa/métodos , Mapeo Restrictivo
13.
Genomics ; 34(1): 97-106, 1996 May 15.
Artículo en Inglés | MEDLINE | ID: mdl-8661029

RESUMEN

We have created a resource for chromosome 22 consisting of 96 unique, well-characterized Fosmids. The Fosmid vector permits efficient cloning of DNA fragments averaging 40 kb in a single-copy vector based on the F factor of Escherichia coli. We have found that Fosmid clones from human chromosome 22 show remarkable stability and are useful for a wide variety of applications in genome analysis. These 96 clones have been localized by FISH, using high-resolution fluorescent banding and multicolor mapping techniques, and their position on the chromosome was correlated with their content of a number of common repeated sequence elements. We identified a subset of clones likely to contain genes by restriction analysis using the enzymes NotI, MluI, SacII, and BssHII. This collection of cytogenetically anchored clones, representing nearly 7% of the chromosome, is of immediate value for detecting chromosomal rearrangements, for use in gene isolation, and as a framework for physical mapping.


Asunto(s)
Mapeo Cromosómico/métodos , Cromosomas Humanos Par 22 , Vectores Genéticos , Sitios de Unión , Centrómero/química , Centrómero/genética , Clonación Molecular , ADN/aislamiento & purificación , ADN Ribosómico/química , ADN Ribosómico/genética , Escherichia coli/genética , Factor F/genética , Humanos , Hibridación Fluorescente in Situ , Secuencias Repetitivas de Ácidos Nucleicos , Mapeo Restrictivo
14.
Nature ; 379(6567): 736-9, 1996 Feb 22.
Artículo en Inglés | MEDLINE | ID: mdl-8602221

RESUMEN

Homozygous staggerer (sg) mice show a characteristic severe cerebellar ataxia due to a cell-autonomous defect in the development of Purkinje cells. These cells show immature morphology, synaptic arrangement, biochemical properties and gene expression, and are reduced in numbers. In addition, sg heterozygotes show accelerated dendritic atrophy and cell loss, suggesting that sg has a role in mature Purkinje cells. Effects of this mutation on cerebellar development have been studied for 25 years, but its molecular basis has remained unknown. We have genetically mapped staggerer to an interval of 160 kilobases on mouse chromosome 9 which was found to contain the gene encoding RORalpha, a member of the nuclear hormone-receptor superfamily. Staggerer mice were found to carry a deletion within the RORalpha gene that prevents translation of the ligand-binding homology domain. We propose a model based on these results, in which RORalpha interacts with the thyroid hormone signalling pathway to induce Purkinje-cell maturation.


Asunto(s)
Proteínas de Unión al ADN/genética , Ratones Mutantes Neurológicos/genética , Receptores de Superficie Celular/genética , Receptores Citoplasmáticos y Nucleares/genética , Transactivadores/genética , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Encéfalo/citología , Encéfalo/embriología , Encéfalo/metabolismo , Diferenciación Celular/fisiología , Ataxia Cerebelosa/genética , Mapeo Cromosómico , ADN Complementario , Proteínas de Unión al ADN/fisiología , Femenino , Masculino , Ratones , Ratones Endogámicos C57BL , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa , Células de Purkinje/patología , Receptores de Superficie Celular/fisiología , Receptores Citoplasmáticos y Nucleares/fisiología , Transactivadores/fisiología
17.
Science ; 270(5244): 1945-54, 1995 Dec 22.
Artículo en Inglés | MEDLINE | ID: mdl-8533086

RESUMEN

A physical map has been constructed of the human genome containing 15,086 sequence-tagged sites (STSs), with an average spacing of 199 kilobases. The project involved assembly of a radiation hybrid map of the human genome containing 6193 loci and incorporated a genetic linkage map of the human genome containing 5264 loci. This information was combined with the results of STS-content screening of 10,850 loci against a yeast artificial chromosome library to produce an integrated map, anchored by the radiation hybrid and genetic maps. The map provides radiation hybrid coverage of 99 percent and physical coverage of 94 percent of the human genome. The map also represents an early step in an international project to generate a transcript map of the human genome, with more than 3235 expressed sequences localized. The STSs in the map provide a scaffold for initiating large-scale sequencing of the human genome.


Asunto(s)
Mapeo Cromosómico , Genoma Humano , Proyecto Genoma Humano , Análisis de Secuencia de ADN , Lugares Marcados de Secuencia , Animales , Línea Celular , Cromosomas Artificiales de Levadura , Bases de Datos Factuales , Expresión Génica , Marcadores Genéticos , Humanos , Células Híbridas , Reacción en Cadena de la Polimerasa
18.
Anal Biochem ; 224(1): 68-74, 1995 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-7710118

RESUMEN

It has been reported that secondary pulsed field gel (SPFG) electrophoresis can dramatically increase the speed of separation of large DNA molecules without a decrease in resolution (Zhang, T. Y., Smith, C. L., and Cantor, C. R. (1991) Nucleic Acids Res. 19, 1291-1296). However, our attempts to duplicate previous SPFG conditions were unsuccessful. We therefore sought to more precisely define the effects of secondary pulsing on the separation of large DNA and to determine the value of the technique in separating molecules up to 1100 kb. Here we report on two of the key SPFG parameters, namely the frequency and duration of the secondary pulse on the migration and resolution of DNA in SPFG. We found that the size range of separation is determined by the sum of the duration of the primary and secondary pulses. Under optimal conditions, a 25-70% increase in velocity can be achieved without loss in resolution.


Asunto(s)
ADN/aislamiento & purificación , Electroforesis en Gel de Campo Pulsado
19.
Nucleic Acids Res ; 18(6): 1481-7, 1990 Mar 25.
Artículo en Inglés | MEDLINE | ID: mdl-2326189

RESUMEN

We have previously shown that asymmetric-voltage field inversion electrophoresis produces more uniform separation for fragments between 1 and 50 kilobases (kb) than other modes of pulsed field gel electrophoresis. We now report on the basis of this phenomenon. As in conventional electrophoresis, the pulsed field mobility of DNAs between 1 and 50 kb varies with voltage in a size dependent manner. The complex migration pattern obtained with asymmetric-voltage field inversion electrophoresis reflects the difference between the mobilities of each sized fragment under the conditions used for the forward and reverse fields. We have applied this technique to DNA sequencing gels and find improvement in resolution for single-stranded fragments in polyacrylamide gels.


Asunto(s)
Secuencia de Bases , ADN/aislamiento & purificación , Electroforesis en Gel de Agar/métodos , Electroforesis/métodos , ADN/genética , ADN de Cadena Simple/genética , ADN de Cadena Simple/aislamiento & purificación , Electroforesis en Gel Bidimensional/métodos , Indicadores y Reactivos , Peso Molecular
20.
Electrophoresis ; 10(5-6): 302-9, 1989.
Artículo en Inglés | MEDLINE | ID: mdl-2527739

RESUMEN

We have studied the migration of DNA in pulsed field agarose gels under a variety of electrophoresis conditions. We have made use of an instrument which can generate electric fields of any orientation, magnitude, or duration to compare different separation techniques for DNA molecules of from 1 to several thousand kilobase pairs. We discuss the capabilities of the system and present results of gel runs in which electrophoresis conditions were changed individually or in combination. The mobility of DNA in pulsed field gels is shown to reflect a number of interdependent physical parameters.


Asunto(s)
ADN/análisis , Electroforesis en Gel de Agar/métodos , Electroforesis/métodos , Autoanálisis/métodos , Bacteriófago lambda/genética , Tampones (Química) , ADN de Hongos/análisis , ADN Viral/análisis , Electrodos , Peso Molecular , Saccharomyces/genética , Temperatura
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