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Recombinant and In Vitro RNA Synthesis

Methods and Protocols

  • Book
  • © 2012

Overview

Editors:
  1. Graeme L. Conn

    1. School of Medicine, Dept. Biochemistry, Emory University, Atlanta, USA

    View editor publications

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  • Details a collection of protocols that guide the reader from the start of a new RNA project to the final sample
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts
  • Includes supplementary material: sn.pub/extras
  • Includes supplementary material: sn.pub/extras

Part of the book series: Methods in Molecular Biology (MIMB, volume 941)

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Table of contents (19 protocols)

  1. Front Matter

    Pages i-xiv
    Download chapter PDF

  2. Purification of RNA Expressed In Vivo Inserted in a tRNA Scaffold

    • Luc Ponchon, Frédéric Dardel
    Pages 1-8

  3. Selective RNase H Cleavage of Target RNAs from a tRNA Scaffold

    • Luc Ponchon, Geneviève Beauvais, Sylvie Nonin-Lecomte, Frédéric Dardel
    Pages 9-18

  4. Preparation of Long Templates for RNA In Vitro Transcription by Recursive PCR

    • Jessica C. Bowman, Bahareh Azizi, Timothy K. Lenz, Poorna Roy, Loren Dean Williams
    Pages 19-41

  5. General Protocols for Preparation of Plasmid DNA Template, RNA In Vitro Transcription, and RNA Purification by Denaturing PAGE

    • Jo L. Linpinsel, Graeme L. Conn
    Pages 43-58

  6. Preparation of Short RNA by In Vitro Transcription

    • Cheng Lu, Pingwei Li
    Pages 59-68

  7. Native RNA Purification by Gel Filtration Chromatography

    • Evan P. Booy, Hui Meng, Sean A. McKenna
    Pages 69-81

  8. Cis-Acting Ribozymes for the Production of RNA In Vitro Transcripts with Defined 5′ and 3′ Ends

    • Johanna M. Avis, Graeme L. Conn, Scott C. Walker
    Pages 83-98

  9. Trans-Acting Antigenomic HDV Ribozyme for Production of In Vitro Transcripts with Homogenous 3′ Ends

    • Milena Szafraniec, Leszek Blaszczyk, Jan Wrzesinski, Jerzy Ciesiolka
    Pages 99-111

  10. Rapid Preparation of RNA Samples Using DNA-Affinity Chromatography and DNAzyme Methods

    • Hae-Kap Cheong, Eunha Hwang, Chaejoon Cheong
    Pages 113-121

  11. Preparation of λN-GST Fusion Protein for Affinity Immobilization of RNA

    • Geneviève Di Tomasso, Philipe Lampron, James G. Omichinski, Pascale Legault
    Pages 123-135

  12. Affinity Purification of RNA Using an ARiBo Tag

    • Geneviève Di Tomasso, Pierre Dagenais, Alexandre Desjardins, Alexis Rompré-Brodeur, Vanessa Delfosse, Pascale Legault
    Pages 137-155

  13. Plasmid Template Design and In Vitro Transcription of Short RNAs Within a “Structure Cassette” for Structure Probing Experiments

    • Virginia K. Vachon, Graeme L. Conn
    Pages 157-169

  14. In Vitro Transcription of Modified RNAs

    • Stephanie L. Moon, Jeffrey Wilusz
    Pages 171-180

  15. End-Labeling Oligonucleotides with Chemical Tags After Synthesis

    • N. Ruth Zearfoss, Sean P. Ryder
    Pages 181-193

  16. High-Purity Enzymatic Synthesis of Site-Specifically Modified tRNA

    • Ya-Ming Hou
    Pages 195-212

  17. Se-Derivatized RNAs for X-ray Crystallography

    • Lina Lin, Zhen Huang
    Pages 213-225

  18. Biosynthetic Preparation of 13C/15N-Labeled rNTPs for High-Resolution NMR Studies of RNAs

    • Luigi Martino, Maria R. Conte
    Pages 227-245

  19. Preparative Separation of Ribonucleoside Monophosphates by Ion-Pair Reverse-Phase HPLC

    • Pierre Dagenais, Pascale Legault
    Pages 247-256

  20. Splint Ligation of RNA with T4 DNA Ligase

    • Christopher J. Kershaw, Raymond T. O’Keefe
    Pages 257-269
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Keywords

About this book

The discovery of catalytic RNAs in the mid-1980s marked the beginning of a new era in RNA biology and an ever increasing appreciation of the diverse and critical roles played by this fascinating molecule. In Recombinant and In Vitro RNA Synthesis: Methods and Protocols, expert researchers in the field detail approaches from the inception of a new RNA project to the final sample ready for analysis. These experimental protocols combine established work-horse approaches with some of the latest innovative methods for RNA synthesis and purification. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and key tips on troubleshooting and avoiding known pitfalls.

 

Authoritative and practical, Recombinant and In Vitro RNA Synthesis: Methods and Protocols aids scientists in continuing to study the fascinating world of RNA.

Editors and Affiliations

  • School of Medicine, Dept. Biochemistry, Emory University, Atlanta, USA

    Graeme L. Conn

Bibliographic Information

  • Book Title: Recombinant and In Vitro RNA Synthesis

  • Book Subtitle: Methods and Protocols

  • Editors: Graeme L. Conn

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-62703-113-4

  • Publisher: Humana Totowa, NJ

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media, LLC 2012

  • Hardcover ISBN: 978-1-62703-112-7Published: 12 October 2012

  • Softcover ISBN: 978-1-4939-5830-6Published: 23 August 2016

  • eBook ISBN: 978-1-62703-113-4Published: 12 October 2012

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 1

  • Number of Pages: XIV, 274

  • Topics: Human Genetics, Nucleic Acid Chemistry

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