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  1. RNA modification
    Autor*in:
    Erschienen: 2007
    Verlag:  Elsevier, Amsterdam

    The presence of modified nucleotides in cellular RNAs has been known for decades and over 100 distinct RNA modifications have been characterized to date. While the exact role of many of these modifications is still unclear, many are highly conserved... mehr

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    The presence of modified nucleotides in cellular RNAs has been known for decades and over 100 distinct RNA modifications have been characterized to date. While the exact role of many of these modifications is still unclear, many are highly conserved across evolution and most contribute to the overall fitness of the organism. In recent years, new methods and bioinformatics approaches have been developed for the dissection of modification pathways and functions. These methods intersect a number of related fields, ranging from RNA processing to comparative genomics and systems biology. In addition, many of the techniques described in this volume have broad applicability, particularly in regards to the isolation, characterization, and reconstitution of ribonucleoprotein complexes, expanding the experimental repertoire available to all RNA researchers

     
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    Hinweise zum Inhalt
    Quelle: Verbundkataloge
    Beteiligt: Gott, Jonatha M. (Hrsg.)
    Sprache: Englisch
    Medientyp: Ebook
    Format: Online
    ISBN: 0123741556
    RVK Klassifikation: WC 4355 ; WC 4460
    Auflage/Ausgabe: Online-Ausg.
    Schriftenreihe: Methods in enzymology ; 425
    Methods in enzymology 0076-6879 ; v. 425
    Schlagworte: RNA editing
    Weitere Schlagworte: RNA Editing
    Umfang: Online-Ressource, illustrations.
    Bemerkung(en):

    Includes bibliographical references and indexes. - Print version record
  2. RNA modification
    Autor*in:
    Erschienen: 2007
    Verlag:  Elsevier, Acad. Press, Amsterdam [u.a.]

    The presence of modified nucleotides in cellular RNAs has been known for decades and over 100 distinct RNA modifications have been characterized to date. While the exact role of many of these modifications is still unclear, many are highly conserved... mehr

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    Universitätsbibliothek Braunschweig
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    Staats- und Universitätsbibliothek Hamburg Carl von Ossietzky
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    The presence of modified nucleotides in cellular RNAs has been known for decades and over 100 distinct RNA modifications have been characterized to date. While the exact role of many of these modifications is still unclear, many are highly conserved across evolution and most contribute to the overall fitness of the organism. In recent years, new methods and bioinformatics approaches have been developed for the dissection of modification pathways and functions. These methods intersect a number of related fields, ranging from RNA processing to comparative genomics and systems biology. In addition, many of the techniques described in this volume have broad applicability, particularly in regards to the isolation, characterization, and reconstitution of ribonucleoprotein complexes, expanding the experimental repertoire available to all RNA researchers.

     
    Export in Literaturverwaltung   RIS-Format
      BibTeX-Format
    Hinweise zum Inhalt
    Quelle: Verbundkataloge
    Beteiligt: Gott, Jonatha M.
    Sprache: Englisch
    Medientyp: Ebook
    Format: Online
    ISBN: 0123741556; 9780123741554
    RVK Klassifikation: VK 8700 ; WC 4355
    Schriftenreihe: Methods in enzymology ; 425
    Schlagworte: RNA editing
    Umfang: Online-Ressource (XLII, 433, [8] S.)
    Bemerkung(en):

    Includes bibliographical references and indexes

    Cover; Contents; Contributors; Preface; Volumes in Series; Section I: Modified Nucleotides; Chapter 1: Identifying Modifications in RNA by MALDI Mass Spectrometry; 1. Introduction; 2. Experimental Strategy; 3. Experimental Procedures; 3.1. Isolation of RNA sequences; 3.2. Digestion of RNA to oligonucleotides; 3.3. MALDI mass spectrometry analysis; 3.4. Tandem mass spectrometry; 3.5. Comparison of MALDI and ESI techniques; 4. Perspectives and Conclusion; Acknowledgments; References; Chapter 2: Identification of Modified Residues in RNAs by Reverse Transcription-Based Methods; 1. Introduction

    2. Reverse Transcription (RT)-Based Methods for Detection of Modified Residues2.1. Reverse transcriptases used for RNA analysis; 2.2. General protocol for primer labeling and reverse transcription of RNA; 3. RNA Extraction from Various Cell Types; 3.1. Protocol for the extraction of total RNA from archaeal or eukaryal cells; 3.2. Protocol for the extraction of total RNA from yeast cells; 4. RNA Modifications Detectable After Specific Chemical Treatment; 4.1. Detection of inosine using glyoxal treatment followed by RNAse T1 hydrolysis; 4.2. Detection of pseudouridine (C) residues

    4.3. Protocol for the detection of pseudouridine residues in RNA using CMCT modification4.4. Complementary approaches for C detection; 4.5. Detection of 5-methylcytosine (m5C); 4.6. Detection of 7-methylguanine (m7G); 4.7. Detection of 20-O-methylated nucleotides using OH cleavage or 20-OH reactivity; 4.8. Protocol for detection of 2'-O-methylated residues in RNA using OH-cleavage; 4.9. Detection of 2'-O-methylated nucleotides by use of low dNTP concentrations; 4.10. Protocol for detection of 2'-O-methylated residues in RNA by use of low dNTP concentrations

    4.11. Other chemical modification methods specific for 20-O-methylations4.12. Detection of dihydrouridine (D) residues by alkaline hydrolysis; 5. RNA Modifications Leading to RT Pauses Without Preliminary Chemical Treatment; 6. Conclusion; References; Section II: tRNA Modifications; Chapter 3: Detection of Enzymatic Activity of Transfer RNA Modification Enzymes Using Radiolabeled tRNA Substrates; 1. Introduction; 2. Identification of Modified Nucleotides in tRNA; 3. Testing the Activity of RNA Modification Enzymes

    3.1. In vitro production of T7-runoff transcripts and nearest-neighbor analysis of modified nucleotides3.2. Experimental conditions for testing activity of tRNA modification enzymes in vitro; 3.3. Optimization of enzymatic reaction conditions; 4. Complete Digestion of RNA with Various Nucleases; 4.1. Use of RNase T2 to generate 3'-monophosphate nucleosides; 4.2. Homemade RNase mix; 4.3. Use nuclease P1 to generate 50-monophosphate nucleosides; 4.4. Use of venom phosphodiesterase VPD, eventually in combination with nuclease P1 to generate 5'-monophosphate nucleosides

    4.5. Use of piperidine to generate mixes of 20- and 30-monophosphate nucleosides

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