Transcription factor protocols / edited by Martin J. Tymms.
Contributor(s): Tymms, Martin JMaterial type: TextSeries: Methods in molecular biology (Clifton, N.J.): v. 130.Publisher: Totowa, N.J. : Humana Press, ©2000Description: 1 online resource (x, 306 pages) : illustrationsContent type: text Media type: computer Carrier type: online resourceISBN: 9781592596867; 159259686X; 1280832878; 9781280832871; 9786610832873; 6610832870Subject(s): Transcription factors -- Laboratory manuals | Transcription Factors -- analysis | Sequence Analysis, DNA -- methods | Transfection -- methods | Facteurs de transcription -- Manuels de laboratoire | SCIENCE -- Life Sciences -- Molecular Biology | Transcription factors | Transfection -- méthodes | Analyse de séquence d'ADN -- méthodes | Facteurs de transcription -- analyse | Méthodes | Wetenschappelijke technieken | Séquençage des acides nucléiques | Facteurs de transcription | Transfection | transcriptiefactoren | transcription factors | transcriptie | transcription | moleculaire biologie | molecular biology | genexpressie | gene expression | polymerase chain reaction | biologische technieken | biological techniques | transcriptomics | protocollen | protocols | Molecular Biology (General) | Transcriptomics | Moleculaire biologie (algemeen)Genre/Form: Electronic books. | Handbooks and manuals. Additional physical formats: Print version:: Transcription factor protocols.DDC classification: 572.8/845 LOC classification: QP552.T68 | T727 2000ebOnline resources: Click here to access online
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Includes bibliographical references and index.
Isolation of target gene promoter/enhancer sequences by whole genome PCR method -- In vivo footprinting using UV light and ligation-mediated PCR -- Identification of DNasel hypersensitive sites within nuclei -- Analysis of in vivo methylation -- Detection of transcription factor partners with a yease one hybrid screen -- Inverse PCR (IPCR) for obtaining promoter sequence -- PCR-directed linker scanning mutagenesis -- Transfection technologies -- The use of particle-mediated gene transfer for the study of promoter activity in somatic tissues -- Optimizing electroporation conditions for the transformation of mammalian cells -- Calcium phosphate transfection of mammalian cultured cells -- DEAE-dextran transfection of mammaliam cultured cells -- Liposome-mediated transfection of mammaliam cells -- Assays for transcriptional activity based on the luciferase reporter gene -- Transient transfection of Schneider cells in the study of transcription factors -- Triplex-forming oligonucleotides and their use in the analysis of gene transcription -- Expression and purification of histidine-tagged transcription factors -- Generation of transcription factors in rabbit reticulocyte lysate depleted of endogenous DNA-binding protein -- Electrophoretic mobility shift assays -- In vitro promoter analysis using nuclear extracts and G-free cassette vectors -- In vitro transcription using competitor oligonucleotides to deplete specific transcription factors -- Computer software of eukaryotic promoter analysis.
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In Transcription Factor Protocols, Martin Tymms has created a powerful compendium of the major techniques for the study of those DNA sequences and protein factors that regulate the transcription of protein encoding genes. With chapters contributed by leading investigators well versed in the methods, this eminently practical compendium includes not only well established protocols, but also novel techniques that are now being widely adopted. Among the important new methods treated are the use of triplex-forming oligonucleotides, the application of whole genome PCR to the isolation of gene promoters/enhancers, the analysis of in vivo methylation, and in vivo footprinting using UV light and ligation-mediated PCR. Step-by-step instructions, tips about pitfalls to avoid, and extensive procedural notes all help to ensure robust and reproducible results. Transcription Factor Protocols offers both experienced workers and new researchers a vital first-stop reference collection of the core techniques for all those exploring the role of transcription factors in gene regulation today.