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Three dimensional imaging of DNA fragments during electrophoresis using a confocal detector

Description: We have measured the three dimensional distribution of DNA fragments within an electrophoretic band. The measurements were made using a confocal microscope and a photon counting photomultiplier detector. A DNA sequencing standard was loaded into glass microchannel plates containing polyacrylamide gel. The measurements were made by scanning the plates in three dimensions using a mechanical stage under computer control, while electrophoresis was taking place. We found that the distribution of DNA was the same for all the bands measured in the sequencing ladder with an approximate Gaussian distribution along all three axes. These measurements are important to understand what physical forces shape electrophoretic bands confined by a channel and also to aid in the design of high throughput DNA sequencers.
Date: January 30, 1995
Creator: Brewer, L.R.; Davidson, C.; Balch, J. & Carrano, A.
Partner: UNT Libraries Government Documents Department

Chromosome 19 International Workshop. Final report, January 1, 1992--December 31, 1992

Description: The Second International Workshop on Human Chromosome 19 was hosted on January 25 and 26, 1992, by the Department of Human Genetics, University Hospital Nijmegen, The Netherlands, at the `Meerdal Conference Center`. The workshop was supported by a grant from the European Community obtained through HUGO, the Dutch Research Organization (NWO) and the Muscular Dystrophy Association (MDA). Travel support for American participants was provided by the Department of Energy. The goals of this workshop were to produce genetic, physical and integrated maps of chromosome 19, to identify inconsistencies and gaps, and to discuss and exchange resources and techniques available for the completion of these maps. The second day of the meeting was largely devoted to region or disease specific efforts. In particular, the meeting served as a platform for assessing and discussing the recent progress made into the molecular elucidation of myotonic dystrophy.
Date: January 4, 1993
Creator: Pericak-Vance, M. A.; Ropers, H. H. & Carrano, A. J.
Partner: UNT Libraries Government Documents Department

A database system for constructing, integrating, and displaying physical maps of chromosome 19

Description: Efforts are underway at numerous sites around the world to construct physical maps of all human chromosomes. These maps will enable researchers to locate, characterize, and eventually understand the genes that control human structure and function. Accomplishing this goal will require a staggering amount of innovation and advancement of biological technology. The volume and complexity of the data already generated requires a sophisticated array of computational support to collect, store, analyze, integrate, and display it in biologically meaningful ways. The Human Genome Center at Livermore has spent the last 6 years constructing a database system to support its physical mapping efforts on human chromosome 19. Our computational support team is composed of experienced computer professionals who share a common pragmatic primary goal of rapidly supplying tools that meet the ever-changing needs of the biologists. Most papers describing computational support of genome research concentrate on mathematical details of key algorithms. However, in this paper we would like to concentrate on the design issues, tradeoffs, and consequences from the point of view of building a complex database system to support leading-edge genomic research. We introduce the topic of physical mapping, discuss the key design issues involved in our databases, and discuss the use of this data by our major tools (DNA fingerprint analysis and overlap computation, contig assembly, map integration, and database browsing.) Given the advantage of hindsight, we discuss what worked, what didn`t, and how we will evolve from here. As early pioneers in this field we hope that our experience may prove useful to others who are now beginning to design and construct similar systems.
Date: June 1, 1994
Creator: Slezak, T.; Wagner, M.; Yeh, Mimi; Ashworth, L.; Nelson, D.; Ow, D. et al.
Partner: UNT Libraries Government Documents Department

Advanced microinstrumentation for rapid DNA sequencing and large DNA fragment separation

Description: Our efforts to develop novel technology for a rapid DNA sequencer and large fragment analysis system based upon gel electrophoresis are described. We are using microfabrication technology to build dense arrays of high speed micro electrophoresis lanes that will ultimately increase the sequencing rate of DNA by at least 100 times the rate of current sequencers. We have demonstrated high resolution DNA fragment separation needed for sequencing in polyacrylamide microgels formed in glass microchannels. We have built prototype arrays of microchannels having up to 48 channels. Significant progress has also been made in developing a sensitive fluorescence detection system based upon a confocal microscope design that will enable the diagnostics and detection of DNA fragments in ultrathin microchannel gels. Development of a rapid DNA sequencer and fragment analysis system will have a major impact on future DNA instrumentation used in clinical, molecular and forensic analysis of DNA fragments.
Date: January 25, 1995
Creator: Balch, J.; Davidson, J.; Brewer, L.; Gingrich, J.; Koo, J.; Mariella, R. et al.
Partner: UNT Libraries Government Documents Department

International workshop of chromosome 19

Description: This document summarizes the workshop on physical and genetic mapping of chromosome 19. The first session discussed the major disease loci found on the chromosome. The second session concentrated on reference families, markers and linkage maps. The third session concentrated on radiation hybrid mapping, somatic cell hybrid panels, macro restriction maps and YACs, followed by cDNA and long range physical maps. The fourth session concentrated on compiling consensus genetic and physical maps as well as discussing regions of conflict. The final session dealt with the LLNL cosmid contig database and comparative mapping of homologous regions of the human and mouse genomes, and ended with a discussion of resource sharing. 18 refs., 2 figs. (MHB)
Date: September 16, 1991
Creator: Pericak-Vance, M.A. (Duke Univ. Medical Center, Durham, NC (United States). Div. of Neurology) & Carrano, A.J. (Lawrence Livermore National Lab., CA (United States))
Partner: UNT Libraries Government Documents Department

Chromosome 19 International Workshop

Description: The Second International Workshop on Human Chromosome 19 was hosted on January 25 and 26, 1992, by the Department of Human Genetics, University Hospital Nijmegen, The Netherlands, at the 'Meerdal Conference Center'. The workshop was supported by a grant from the European Community obtained through HUGO, the Dutch Research Organization (NWO) and the Muscular Dystrophy Association (MDA). Travel support for American participants was provided by the Department of Energy. The goals of this workshop were to produce genetic, physical and integrated maps of chromosome 19, to identify inconsistencies and gaps, and to discuss and exchange resources and techniques available for the completion of these maps. The second day of the meeting was largely devoted to region or disease specific efforts. In particular, the meeting served as a platform for assessing and discussing the recent progress made into the molecular elucidation of myotonic dystrophy.
Date: January 4, 1993
Creator: Pericak-Vance, M.A. (Duke Univ., Durham, NC (United States). Medical Center); Ropers, H.H. (Univ. Hospital Nijmegen, (The Netherlands). Dept. of Human Genetics) & Carrano, A.J. (Lawrence Livermore National Lab., CA (United States))
Partner: UNT Libraries Government Documents Department

The construction of a physical map for human chromosome 19

Description: Unlike a genetic map which provides information on the relative position of genes or markers based upon the frequency of genetic recombination, a physical map provides a topographical picture of DNA, i.e. distances in base pairs between landmarks. The landmarks may be genes, gene markers, anonymous sequences, or cloned DNA fragments. Perhaps the most useful type of physical map is one that consists of an overlapping set of cloned DNA fragments (contigs) that span the chromosome. Once genes are assigned to this contig map, sequencing of the genomic DNA can be prioritized to complete the most interesting regions first. While, in practice, complete coverage of a complex genome in recombinant clones may not be possible to achieve, many gaps in a clone map may be closed by using multiple cloning vectors or uncloned large DNA fragments such as those separated by electrophoretic methods. Human chromosome 19 contains about 60 million base pairs of DNA and represents about 2% of the haploid genome. Our initial interest in chromosome 19 originated from the presence of three DNA repair genes which we localized to a region of this chromosome. Our approach to constructing a physical map of human chromosome 19 involves four steps: building a foundation of overlapping cosmid contigs; bridging the gaps in the cosmid map with hybridization-based methods to walk onto DNA cloned in yeast and cosmids: orienting the contigs relative to each other and linking them to the cytological map; and coupling the contig map with the genetic map. The methods we use and the current status of the map are summarized in this report. 6 refs., 1 fig.
Date: November 5, 1990
Creator: Carrano, A.V.; Alleman, J.; Amemiya, C.; Ashworth, L.K.; Aslanidis, C.; Branscomb, E.W. et al.
Partner: UNT Libraries Government Documents Department