&i;Circadian Rhythms: Methods and Protocols&o; is an in-depth examination of circadian biology, presented by leading researchers in the field. This cutting-edge collection of protocols is adaptable for research at every level, and represents the huge strides that chronobiologists have made over the past two decades. These robust, reproducible procedures take into account the myriad of variables that circadian researchers must be cognizant of while conducting their research.
By distilling the arsenal of methodologies available, and presenting essential procedures in precise detail, this book supports a vast majority of circadian research efforts.
Part I of this seven-part volume provides a concise introduction to essential components of circadian biology, including light, photoreceptors, circadian clocks, and statistical analysis. Part II presents protocols and techniques for monitoring and analyzing rhythmic patterns of several model organisms. Parts III, IV, and V emphasize the identification of clock genes and protein and RNA expression in circadian biology. Protocols for in vitro systems comprise Part VI, and Part VII provides an outline of methods and tips for microscopy analysis.
Circadian biologists at all research levels will realize tremendous benefit from this extraordinary collection of methods and protocols for the study of circadian rhythms.
Preface; Contributors; I. Overviews.; 1. Light, photoreceptors and circadian clocks; Russell G. Foster, Mark W. Hankins and Stuart N. Peirson; 2. Statistical analysis of biological rhythm data; Harold B. Dowse; II. Rhythmic Readouts.; 3. Rhythmic conidiation in Neurospora crassa.; Cas Kramer; 4. Monitoring and analyzing Drosophila circadian locomotor activity.; Mauro A. Zordan, Clara Benna and Gabriella Mazzotta.; 5. Automated video image analysis of larval zebrafish locomotor rhythms.; Gregory M. Cahill; 6. Locomotor activity in rodents.; Gianluca Tosini; 7. Analysis of circadian leaf movement rhythms in Arabidopsis thaliana.; Kieron D. Edwards and Andrew J. Millar; 8. Detection of rhythmic bioluminescence from luciferase reporters in cyanobacteria.; Shannon R. Canales, Jayna L. Ditty, Eugenia M. Clerico and Susan S. Golden; 9. Analysis of rhythmic gene expression in adult Drosophila using the firefly-luciferase reporter gene.; Ralf Stanewsky; 10. Monitoring circadian rhythms in Arabidopsis thaliana using luciferase reporter genes.; Anthony Hall and Paul Brown; III. Identification of Clock Genes.; 11. Specialized techniques for site directed mutagenesis in cyanobacteria; Eugenia M. Clerico, Jayna L. Ditty and Susan S. Golden; 12. Novel strategies for the identification of clock genes in Neurospora with insertional mutagenesis; Kruno Sveric, Moyra Mason, Till Roenneberg and Martha Merrow; 13. Mutagenesis with Drosophila; Patrick Emery; 14. Mutagenesis in Arabidopsis.; Jodi Maple and Simon G. Moller; 15. Yeast-two-hybrid screening. Jodi Maple and Simon G. Moller; 16. Microarrays: quality control and hybridization protocol.; Ken-ichiro Uno and Hiroki R. Ueda; 17. Microarrays: statistical methods for circadian rhythms.; Rikuhiro Yamada and Hiroki R. Ueda; 18. Identification of clock genes using difference gel electrophoresis (DIGE).; Natasha A. Karp and Kathryn S. Lilley; IV. Gene Expression: RNA.; 19. Isolation of total RNA from Neurospora mycelium.; Cas Kramer; 20. RNA extraction from Drosophila heads.; Patrick Emery; 21. Extraction of plant RNA.; Michael G. Salter and Helen E. Conlon; 22. RNA Extraction from mammalian tissues.; Stuart N. Peirson and Jason N. Butler; 23. Northern analysis of sense and antisense frequency RNA in Neurospora crassa.; Cas Kramer; 24. RNase protection assay.; Patrick Emery; 25. Quantitative PCR.; Stuart N. Peirson and Jason N. Butler; V. Gene Expression: Proteins.; 26. Protein extraction, fractionation and purification from cyanobacteria.; Natalia B. Ivleva and Susan S. Golden; 27. Protein extraction from Drosophila heads.; Patrick Emery; 28. Plant protein extraction.; Helen E. Conlon and Michael G. Salter; 29. Protein extraction from mammalian tissues.; Choogon Lee.