This volume contains state-of-the-art methods tackling all aspects of small non-coding RNAs biology.Small Non-Coding RNAs: Methods and Protocols guides readers through customized dedicated protocols and technologies that will be of valuable help to all those willing to contribute deciphering the numerous functions of small non-coding RNAs. Written in the highly successful Methods of 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 troubles troubleshooting and avoiding known pitfalls.
Instructive and practical, Small Non-Coding RNAs: Methods and Protocols reaches out to biochemists, cellular, and molecular biologists already working in the field of RNA biology and to those just starting to study small non-coding RNAs.
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Part I Introduction to Small Non-Coding RNAs |
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1 Small Non-Coding RNAs: A Quick Look in the Rearview Mirror |
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3 | (8) |
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2 Alcoholic Precipitation of Small Non-Coding RNAs |
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11 | (6) |
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3 Quantification and Quality Control of a Small Non-Coding RNA Preparation |
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17 | (12) |
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4 Impact of RNA Isolation Protocols on RNA Detection by Northern Blotting |
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29 | (12) |
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Part II Visualization and Analysis of Small Non-Coding RNAs |
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5 Improved Northern Blot Detection of Small RNAs Using EDC Crosslinking and DNA/LNA Probes |
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41 | (12) |
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6 Direct Cloning of Double-Stranded RNAs |
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53 | (12) |
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7 Detection and Labeling of Small Non-Coding RNAs by Splinted Ligation |
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65 | (8) |
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8 Fluorescence In Situ Hybridization of Small Non-Coding RNAs |
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73 | (12) |
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9 RT-qPCR-Based Quantification of Small Non-Coding RNAs |
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85 | (18) |
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10 Stem-Loop RT-PCR Based Quantification of Small Non-Coding RNAs |
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103 | (6) |
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11 miR-RACE: An Effective Approach to Accurately Determine the Sequence of Computationally Identified miRNAs |
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109 | (10) |
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12 Probing Small Non-Coding RNAs Structures |
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119 | (20) |
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Part III High-Throughput Approaches to Study Non-Coding RNAs |
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13 cDNA Library Generation for the Analysis of Small RNAs by High-Throughput Sequencing |
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139 | (12) |
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14 CLIP-Seq to Discover Transcriptome-Wide Imprinting of RNA Binding Proteins in Living Cells |
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151 | (10) |
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15 Microarray Analysis of Small Non-Coding RNAs |
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161 | (14) |
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Part IV Small Non-Coding RNAs Applications |
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16 RLM-RACE, PPM-RACE, and qRT-PCR: An Integrated Strategy to Accurately Validate miRNA Target Genes |
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175 | (12) |
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17 Dual Luciferase Gene Reporter Assays to Study miRNA Function |
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187 | (12) |
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18 Gene Expression Knockdown by Transfection of siRNAs into Mammalian Cells |
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199 | (4) |
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19 Efficient and Selective Knockdown of Small Non-Coding RNAs |
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203 | (10) |
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20 Cell-SELEX: In Vitro Selection of Synthetic Small Specific Ligands |
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213 | (12) |
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21 Small Non-Coding RNAs and Aptamers in Diagnostics and Therapeutics |
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225 | (10) |
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| Index |
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Mathieu Rederstorff, PhD Associate Professor Laboratoire IMoPA UMR 7365 CNRS/UL Equipe ARN, RNP - Maturation, Structure, Fonction - Université de Lorraine Biopôle/Campus Médecine Brabois 9 Avenue de la Forêt de Haye BP 184 54506 Vandoeuvre-lès-Nancy FRANCE Tel: +33 3 83 68 55 31 (office) +33 3 83 68 55 25 (lab) +33 6 49 12 67 11 (mobile) Fax: +33 3 83 68 55 09 (fax) mathieu.rederstorff@univ-lorraine.fr mathieu.rederstorff@maem.uhp-nancy.fr
Lisainfo e-raamatute kohta