RNA Methodologies : A Laboratory Guide for Isolation and Characterization
This laboratory guide is a collection of tested and optimized laboratory protocols for the isolation and characterization of eukaryotic RNA. This edition includes a variety of RNA-based PCR techniques, such as differential display PCR, and transcript quantification by competitive (mimic) PCR. The book focuses on how these methodologies can be used in the study of transcriptional and posttranscriptional regulation of eukaryotic genes. The liberal incorporation of flow charts, tables, and graphs facilitate learning and assist in the planning phases of projects. It is a valuable resource for investigators who wish to characterize cellular biochemistry.
- Paperback | 533 pages
- 167.9 x 227.6 x 24.4mm | 821.12g
- 01 Oct 1998
- Elsevier Science Publishing Co Inc
- Academic Press Inc
- San Diego, United States
- 2nd Revised edition
- b&w illustrations, charts, graphs, tables
Each of the protocols is written relatively independent of the rest of the text, allowing the reader to immediately begin using a procedure without having to study the text from cover to cover... Very few other molecular biology guide books are available that cover the isolation and manipulation of RNA as exhaustingly as does this one. This book would be highly valuable in any research facility engaged in molecular biology. --Mark A. Jandreski, Loyola University Medical Center, DOODY'S ELECTRONIC JOURNAL
Table of contents
Preface. RNA and the Cellular Biochemistry Revisited. Transcription and the Organization of Eukaryotic Gene Sequences. Messenger RNA. Resilient Ribonucleases. RNA Isolation Strategies. Isolation of Polyadenylated RNA. Quality Control for RNA Preparations. Dot Blot Analysis. Electrophoresis of RNA. Photodocumentation and Image Analysis. The Northern Analysis. Nucleic Acid Probe Technology. Practical Nucleic Acid Hybridization. Principles of Detection. RT PCR. Messenger RNA Differential Display. Quantitative PCR Techniques. Quantification of Specific mRNAs by Nuclease Protection. Analysis of Nuclear RNA. An RNA Paradigm. Epilogue. A Few Pearls of Wisdom. Appendices: Phenol Preparation. Disposal of Ethidium Bromide and SYBR Green Solutions. DNase I Removal of DNA from an RNA Sample. RNase Incubation to Remove RNA from a DNA sample. Deionization of Formamide, Formaldehyde, and Glyoxal. Silanizing Centrifuge Tubes and Glassware. Centrifugation as a Mainstream Tool for the Molecular Biologist. Trypsinization Protocol for Anchorage-Dependent Cells. Isolation of High-Molecular-Weight DNA by the Salting-Out Procedure. RNA Isolation from Plant Tissue. Electrophoresis: Principles, Parameters, and Safety. Polyacrylamide Gels. Internal Controls. Trade Citations. Selected Suppliers of Equipment, Reagents, and Services. Glossary. Index.