RNAi and plant gene function analysis : methods and protocols
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Bibliographic Information
RNAi and plant gene function analysis : methods and protocols
(Springer protocols)(Methods in molecular biology / John M. Walker, series editor, v. 744)
Humana Press, c2011
- : hbk
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Includes bibliographical references and index
Description and Table of Contents
Description
The use of RNAi technology is essential for most plant science researchers. As DNA sequence information increases, so the need for functional annotation of target genes also increases. Authoritative and accessible, RNAi and Plant Gene Function Analysis: Methods and Protocols seeks to serve as an ideal guide to scientists of all backgrounds looking to further their knowledge of this fundamental science. Written in the highly successful Methods in Molecular Biology (TM) series format, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and notes on troubleshooting and avoiding known pitfalls.
In addition to the functional annotation of unknown genes, RNAi technology has been applied to the genetic engineering of important plant metabolites including starches, oils, and storage proteins. RNAi has been used to engineer plants resistant to plant viruses and also to nematodes and insects. RNAi and Plant Gene Function Analysis: Methods and Protocols will provide both professionals and novices of plant biotechnology and breeding research with essential tips for designing experiments meant to explore plant gene function.
Table of Contents
1. Overview of Plant RNAi
Yuichiro Watanabe
2. Caveat of RNAi in Plants: the Off-Target Effect
Muthappa Senthil-Kumar and Kirankumar S. Mysore
3. Plant Gateway Vectors for RNAi as a Tool for Functional Genomic Studies
Toshiya Muranaka
4. Heat-Inducible RNAi for Gene Functional Analysis in Plants
Frederic Masclaux and Jean-Philippe Galaud
5. Gene Function Analysis by Artificial MicroRNAs in Physcomitrella patens
Basel Khraiwesh, Isam Fattash, M. Asif Arif, and Wolfgang Frank
6. Virus-Induced Gene Silencing in Ornamental Plants
Cai-Zhong Jiang, Jen-Chih Chen, and Michael Reid
7. Local RNA Silencing Mediated by Agroinfiltration
Jutta Maria Helm, Elena Dadami, and Kriton Kalantidis
8. Direct Transfer of Synthetic Double-Stranded RNA into Protoplasts of Arabidopsis thaliana
Ha-il Jung, Zhiyang Zhai and Olena K. Vatamaniuk
9. Detection of Long and Short Double-Stranded RNAs
Toshiyuki Fukuhara, Syunichi Urayama, Ryo Okada, Eri Kiyota, and Hiromitsu Moriyama
10. Quantitative Stem-Loop RT-PCR for Detection of MicroRNAs
Erika Varkonyi-Gasic and Roger P. Hellens
11. Large Scale Sequencing of Plant Small RNAs
William P. Donovan, Yuanji Zhang, and Miya D. Howell
12. Computational Prediction of Plant miRNA Targets
Ying-Hsuan Sun, Shanfa Lu, Rui Shi, and Vincent L. Chiang
13. Bisulfite Sequencing for Cytosine-Methylation Analysis in Plants
Nazmul Haque and Masamichi Nishiguchi
14. Using Nuclear Run-On Transcription Assays in RNAi Studies
Basel Khraiwesh
15. Proteomic Analysis of RNA-Interference Induced Knockdown Plant
Sang Yeol Lee and Kyun Oh Lee
16. Comparative Analysis of Phosphoprotein Expression using 2D-DIGE
Tomoya Asano and Takumi Nishiuchi
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