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Chromosome Specific Cdnas

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Chromosome Specific Cdnas

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Author :
language : en
Publisher:
Release Date : 1991

Chromosome Specific Cdnas written by and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1991 with categories.

This project has three main goals: (1) to construct high-quality normalized cDNA libraries from human tissues, (2) to develop methods for isolation of chromosome-specific cDNAs, (3) to contribute sequence information to the expanding cDNA/EST database. Our short term goal has been to construct a very high quality cDNA library from a 3 month post-natal total human brain. Several libraries were constructed and carefully characterized by DNA sequencing and restriction analysis while protocols were being streamlined. We have now generated a library which has the following main features: (1) very high complexity (107 recombinants), (b) short poly (A) tails; (c) long size inserts (1 kb average); (d) undetectable co-cloning events; (e) background of non-recombinants. The next step will be to generate and test a normalized version of this brain cDNA library. Subsequently, single-stranded versions of the normalized library will be utilized for isolation of chromosome 13-specific cDNAs.

Chromosome Specific Cdnas Stss Progress Report September 1 1991 February 28 1992

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Author :
language : en
Publisher:
Release Date : 1991

Chromosome Specific Cdnas Stss Progress Report September 1 1991 February 28 1992 written by and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1991 with categories.

Positional Cloning By Exon Trapping And Cdna Selection

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Author : Bernhard Korn
language : en
Publisher: John Wiley & Sons
Release Date : 1999-06-02

Positional Cloning By Exon Trapping And Cdna Selection written by Bernhard Korn and has been published by John Wiley & Sons this book supported file pdf, txt, epub, kindle and other format this book has been release on 1999-06-02 with Science categories.

Tremendous strides in decoding the human genome have been made over the last ten years. Large, chromosome-scale, genome-scale EST-sequencing and mapping techniques have dramatically expanded the base of identified genes and sequenced DNA. Gene isolation and cloning techniques that emerged earlier on, however, still remain effective mainstays for constructing dense transcript maps to isolate coding sequences and disease-associated genes contained within a specific chromosomal region. Positional Cloning by Exon Trapping and cDNA Selection examines two powerful methods for locating the coding region of a given gene by isolating gene fragments from individual clones or pools of genomic clones. This comprehensive guide details the exon-trapping and cDNA selection processes step by step-from isolating genomic templates and nuclear splicing, to verifying generated clones and sequenced data, to analyzing exon libraries and cDNA sublibraries. Procedures covered include: * Exon-trapping systems and descriptions of pSPL1 and pSPL3 vectors. * Exon amplification protocols for preparing vectors for cloning; subcloning genomic DNA into vectors; transformation, analysis, and transfection of sublibraries; RNA transcription; and PCR amplification and PCR product cloning. * Evaluation of exon libraries by PCR colony testing, identifying artifactual clones using Southern blot, and sequencing and mapping back candidate exons. * Isolation of genomic templates, such as COSMID-, P1-, PAC, and YAC DNA. * cDNA selection experiment protocols including cDNA screening, hybridization, and biotinylation; preparation of genomic and cDNA sources; and PCR cloning. * Clone analysis and analysis by hybridization. Positional Cloning by Exon Trapping and cDNA Selection offers researchers, scientists, and graduate students an invaluable tool for probing gene distribution and molecular organization. Most importantly, it provides a critical approach to isolating specific disease genes within a targeted genomic area.

An Improved Methodology For Identifying Chromosome Specific Genes From A Cdna Library

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Author : Nancy Soo Hyung Bae
language : en
Publisher:
Release Date : 1995

An Improved Methodology For Identifying Chromosome Specific Genes From A Cdna Library written by Nancy Soo Hyung Bae and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1995 with Gene libraries categories.

Cloning Human Chromosome 17 Genes

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Author :
language : en
Publisher:
Release Date : 1995

Cloning Human Chromosome 17 Genes written by and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1995 with categories.

Our research interest is focused on the development of new strategies to identify genes from chromosome 17. The isolation of genes transcribed from chromosome 17 will provide candidates for the proposed sporadic breast and ovarian cancer genes. We have recently reported a method for the isolation of chromosome specific cDNAs using high density arrayed cDNA and chromosome specific cosmid libraries. The ability to isolate genes in a chromosome specific manner provides simultaneous identification of the expressed sequence and a chromosomal location. This new technology identifies expressed sequences by reciprocal probing of arrayed cDNA libraries and a chromosome specific cosmid library. We have used these resources to identify 1794 clones from the Los Mamos chromosome 17 cosmid library using probes generated from a placental cDNA library. So far, we have isolated and characterized 42 cDNAs to chromosome 17. Several of these cDNAs mapped to the region 17p13, while 14 other cDNAs mapped to the region of 17q12-22. Recent studies have demonstrated that in some sporadic breast cancer there are loss of heterozygosity (LOH) in these regions of human chromosome 17.

Chromosome Microdissection And Cloning

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Author : Nabil Hagag
language : en
Publisher: Academic Press
Release Date : 2012-12-02

Chromosome Microdissection And Cloning written by Nabil Hagag and has been published by Academic Press this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-12-02 with Science categories.

Chromosome Microdissection and Cloning: A Practical Guide is a straightforward guide to chromosome microdissection and cloning. It presents an overview of the procedures and briefly reviews a few areas of research in which these techniques are applied. Topics range from preparation of chromosomes for microdissection to molecular cloning of microdissected chromosomal DNA. Methods of chromosome microdissection, including video microscope method and oil chamber method, are described. Comprised of five chapters, this book begins with an overview of the structure and organization of chromosomes, followed by a description of methods for preparing and preserving chromosomal DNA in a manner that is useful for cloning and direct analysis. Microdissection of metaphase chromosomes and isolation of fragments can be accomplished in one of the three ways described in the next chapter: by microdissection using an upright microscope and glass capillaries in an oil chamber; by laser microbeam; and with the use of an inverted microscope equipped with a video camera and high magnification-high resolution lenses. A step-by-step guide to these techniques and solutions for common problems are given following each method. Protocols for cloning and identifying genetic sequences from defined chromosome regions, particularly using the polymerase chain reaction, are also discussed. The final chapter focuses on applications of chromosome microdissection, such as cloning of disease-specific genes and generating "sequence tagged sites" to be used in large DNA sequencing projects. This monograph will be particularly helpful to investigators setting up microdissection systems de novo.

Human Chromosome 21

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Author :
language : en
Publisher:
Release Date : 1991

Human Chromosome 21 written by and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1991 with categories.

The objective of the research funded by DOE grant DE-FG02-89ER60857 from 6/15/89 to 8/31/91 was to contribute to the physical mapping of human chromosome 21 (HC21) by cloning large fragments of DNA into Yeast Artificial Chromosomes (YACs) and identify YACs that map on HC21. A total of 54 sequence tagged sites (STS) have been developed and mapped in our laboratory to HC21 and can be used as initial reference points for YAC identification and construction of overlapping clones. A small YAC library was constructed which is HC21 specific. DNA from somatic cell hybrid WAV17 or from flow-sorted HC21 was partially digested with EcoRI, ligated into vectors PJS97, PJS98, and YACs have been obtained with average size insert of more than 300 kb. This library has been deposited in D. Patterson's lab for the Joint YAC screening effort. Additional YAC libraries from ICI Pharmaceuticals or from Los Alamos National Laboratories have been screened with several STS and positive YACs have been identified. Work in progress includes screening of YAC libraries in order to construct overlapping clones, characterization of the cloning ends of YACs, characterization of additional STS and cloning of HC21 specific cDNAs. 15 refs., 2 figs., 5 tabs.

Identification Of Transcribed Sequences

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Author : K. Gardiner
language : en
Publisher: Springer Science & Business Media
Release Date : 2012-12-06

Identification Of Transcribed Sequences written by K. Gardiner and has been published by Springer Science & Business Media this book supported file pdf, txt, epub, kindle and other format this book has been release on 2012-12-06 with Science categories.

The Human Genome Project, an endeavor to map and sequence the entire human genome, has been in existence for almost seven years. One result of this effort has been the development of increasingly detailed genetic and physical maps spanning large regions of virtually every chromosome. Paralleling this has been the increasingly high resolution mapping of many &wnetic diseases. Together, these developments have facilitated the isolation of specific disease genes and are now motivating the construction of comprehensive transcriptional maps. This latter endeavor represents a new facet of the genome project, and as such requires the recognition and solution of a new set of problems, with the attendant development and application of a new set of techniques. The First International Workshop on the Identification of Transcribed Sequences in the Human Genome was held in 1991 and was attended by 23 investigators. Discussions at this meeting were largely devoted to defining the magnitude of the problem and describing the available techniques. A small number of laboratories reported the development of new techniques (at that time, for example, exon trapping, cDNA hybrid selection, direct cDNA screening, use of splice junction conserved sequences,etc.), but data were too limited to permit comparisons of their relative efficiencies.

Cloning Human Chromosome 17 Genes Candidate Genes For Brcal

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Author :
language : en
Publisher:
Release Date : 1996

Cloning Human Chromosome 17 Genes Candidate Genes For Brcal written by and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1996 with categories.

From loss of heterozygosity (LOH) studies, it has been determined that three regions 17p13, 17q12-22, and 17q24-25 of human chromosome 17 are frequently deleted in breast cancer. The presence of LOH in these region suggest the location of putative tumor suppressor. Our research is focused on the isolation of chromosome 17 genes. A method for the isolation of chromosome specific cDNAs using high density arrayed cDNA and chromosome specific cosmid libraries was developed. To date we have isolated and mapped 105 unique cDNAs of chromosome 17. Of these we have mapped 72 of these cDNAs to the three regions that have been determined to be associated with LOH in breast cancer. These cDNAs are now potential candidate genes for the putative tumor suppressor associated with breast cancer on human chromosome 17. Potential function of these cDNAs will be determined by comparing sequence information to known protein motif in the genome data base. Genes encoding for proteins involved in cellular functions including signal transduction, transcription and cell cycle pathways are prime candidate for tumor suppressor. These genes will be further characterize to determine if they play any role in breast cancer etiology.

Isolation Of Cdnas From The Human X Chromosome And Derivation Of Related Stss Final Progress Report April 1992 March 1995

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Author :
language : en
Publisher:
Release Date : 1995

Isolation Of Cdnas From The Human X Chromosome And Derivation Of Related Stss Final Progress Report April 1992 March 1995 written by and has been published by this book supported file pdf, txt, epub, kindle and other format this book has been release on 1995 with categories.

Over the course of this funding period, the number of genes assigned to the human X chromosome has approximately tripled from less than one hundred to nearly three hundred characterized, cloned genes assigned to it. The aims of this project were to develop methods for gene identification and to identify and characterize expressed sequences from the X chromosome. The rapidly changing environment of the human genome project provided abundant resources for gene characterization, and since methods for gene identification became rather robust over this period, these aims were de-emphasized during the project. Among the methods developed was a local one (reciprocal probing) that was developed by Drs. Cheng Chi Lee and C. Thomas Caskey, with emphasis on the human X chromosome. The development of this method offered significant expressed sequence resources for this project, particularly when coupled with the efforts to identify cosmid clones from specific X chromosome locations, as the reciprocal probing process results in paired genomic (cosmid) and cDNA materials. Attention, then has been paid to characterization of genes rather than to their identification.

Chromosome Specific Cdnas

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