Recombinant and in vitro RNA synthesis : methods and protocols / edited by Graeme L. Conn.

Contributor(s): Conn, Graeme L [edt]
Material type: TextTextSeries: Methods in molecular biology (Clifton, N.J.): v. 941.Publisher: New York : Humana Press : Springer, ©2012Description: 1 online resource (xiv, 274 pages) : illustrationsContent type: text Media type: computer Carrier type: online resourceISBN: 9781627031134; 1627031138Subject(s): RNA | RNA splicing | RNA | RNA Splicing | RNA | RNA splicing | biomedische wetenschappen | biomedicine | nucleic acids | chemie | chemistry | humane genetica | human genetics | Medicine (General) | Geneeskunde (algemeen)Genre/Form: Electronic books. Additional physical formats: Print version:: Recombinant and in vitro RNA synthesis.DDC classification: 611.01816 LOC classification: RB155-155.8Online resources: Click here to access online
Contents:
Purification of RNA expressed in vivo inserted in a tRNA scaffold / Luc Ponchon and Frederic Dardel -- Selective RNase H cleavage of target RNAs from a tRNA scaffold / Luc Ponchon [and others] -- Preparation of long templates for RNA in vitro transcription by recursive PCR / Jessica C. Bowman [and others] -- General protocols for preparation of plasmid DNA template, RNA in vitro transcription, and RNA purification by denaturing PAGE / Jo L. Linpinsel and Graeme L. Conn -- Preparation of short RNA by in vitro transcription / Cheng Lu and Pingwei Li -- Native RNA purification by gel filtration chromatography / Evan P. Booy, Hui Meng, and Sean A. McKenna -- Cis-acting ribozymes for the production of RNA in vitro transcripts with defined 5ʹ and 3ʹ ends / Johanna M. Avis, Graeme L. Conn, and Scott C. Walker -- Trans-acting antigenomic HDV ribozyme for production of in vitro transcripts with homogenous 3ʹ ends / Milena Szafraniec [and others] -- Rapid preparation of RNA samples using DNA-affinity chromatography and DNAzyme methods / Hae-Kap Cheong, Eunha Hwang, and Chaejoon Cheong -- Preparation of lN-GST fusion protein for affinity immobilization of RNA / Genevieve Di Tomasso [and others] -- Affinity purification of RNA using an ARiBo tag / Genevieve Di Tomasso [and others] -- Plasmid template design and in vitro transcription of short RNAs within a "structure cassette" for structure probing experiments / Virginia K. Vachon and Graeme L. Conn -- In vitro transcription of modified RNAs / Stephanie L. Moon and Jeffrey Wilusz -- End-labeling oligonucleotides with chemical tags after synthesis / N. Ruth Zearfoss and Sean P. Ryder -- High-purity enzymatic synthesis of site-specifically modified tRNA / Ya-Ming Hou -- Se-derivatized RNAs for x-ray crystallography / Lina Lin and Zhen Huang -- Biosynthetic preparation of 13C/15N-labeled rNTPs for high-resolution NMR studies of RNAs / Luigi Martino and Maria R. Conte -- Preparative separation of ribonucleoside monophosphates by ion-pair reverse-phase HPLC / Pierre Dagenais and Pascale Legault -- Splint ligation of RNA with T4 DNA ligase / Christopher J. Kershaw and Raymond T. O'Keefe.
Summary: 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.
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Purification of RNA expressed in vivo inserted in a tRNA scaffold / Luc Ponchon and Frederic Dardel -- Selective RNase H cleavage of target RNAs from a tRNA scaffold / Luc Ponchon [and others] -- Preparation of long templates for RNA in vitro transcription by recursive PCR / Jessica C. Bowman [and others] -- General protocols for preparation of plasmid DNA template, RNA in vitro transcription, and RNA purification by denaturing PAGE / Jo L. Linpinsel and Graeme L. Conn -- Preparation of short RNA by in vitro transcription / Cheng Lu and Pingwei Li -- Native RNA purification by gel filtration chromatography / Evan P. Booy, Hui Meng, and Sean A. McKenna -- Cis-acting ribozymes for the production of RNA in vitro transcripts with defined 5ʹ and 3ʹ ends / Johanna M. Avis, Graeme L. Conn, and Scott C. Walker -- Trans-acting antigenomic HDV ribozyme for production of in vitro transcripts with homogenous 3ʹ ends / Milena Szafraniec [and others] -- Rapid preparation of RNA samples using DNA-affinity chromatography and DNAzyme methods / Hae-Kap Cheong, Eunha Hwang, and Chaejoon Cheong -- Preparation of lN-GST fusion protein for affinity immobilization of RNA / Genevieve Di Tomasso [and others] -- Affinity purification of RNA using an ARiBo tag / Genevieve Di Tomasso [and others] -- Plasmid template design and in vitro transcription of short RNAs within a "structure cassette" for structure probing experiments / Virginia K. Vachon and Graeme L. Conn -- In vitro transcription of modified RNAs / Stephanie L. Moon and Jeffrey Wilusz -- End-labeling oligonucleotides with chemical tags after synthesis / N. Ruth Zearfoss and Sean P. Ryder -- High-purity enzymatic synthesis of site-specifically modified tRNA / Ya-Ming Hou -- Se-derivatized RNAs for x-ray crystallography / Lina Lin and Zhen Huang -- Biosynthetic preparation of 13C/15N-labeled rNTPs for high-resolution NMR studies of RNAs / Luigi Martino and Maria R. Conte -- Preparative separation of ribonucleoside monophosphates by ion-pair reverse-phase HPLC / Pierre Dagenais and Pascale Legault -- Splint ligation of RNA with T4 DNA ligase / Christopher J. Kershaw and Raymond T. O'Keefe.

Includes bibliographical references and index.

Print version record.

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.

English.

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