Conn G. (Ed.) Recombinant and In Vitro RNA Synthesis: Methods and Protocols

Humana Press, 2012. - 278 p. ISBN: 162703112XThe 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.Purification of RNA Expressed In Vivo Inserted in a tRNA Scaffold
Selective RNase H Cleavage of Target RNAs from a tRNA Scaffold
Preparation of Long Templates for RNA In Vitro Transcription by Recursive PCR
General Protocols for Preparation of Plasmid DNA Template, RNA In Vitro Transcription, and RNA Purification by Denaturing PAGE
Preparation of Short RNA by In Vitro Transcription
Native RNA Purification by Gel Filtration Chromatography
Cis-Acting Ribozymes for the Production of RNA In Vitro Transcripts with Defined 5′ and 3′ Ends
Trans-Acting Antigenomic HDV Ribozyme for Production of In Vitro Transcripts with Homogenous 3′ Ends
Rapid Preparation of RNA Samples Using DNA-Affinity Chromatography and DNAzyme Methods
Preparation of lN-GST Fusion Protein for Affinity Immobilization of RNA
Affinity Purification of RNA Using an ARiBo Tag
Plasmid Template Design and In Vitro Transcription of Short RNAs Within a Structure Cassette for Structure Probing Experiments
In Vitro Transcription of Modified RNAs
End-Labeling Oligonucleotides with Chemical Tags After Synthesis
High-Purity Enzymatic Synthesis of Site-Specifically Modified tRNA
Se-Derivatized RNAs for X-ray Crystallography
Biosynthetic Preparation of 13C/15N-Labeled rNTPs for High-Resolution NMR Studies of RNAs
Preparative Separation of Ribonucleoside Monophosphates by Ion-Pair Reverse-Phase HPLC
Splint Ligation of RNA with T4 DNA Ligase