Captures the current state of this ever-developing and powerful technology
Features practical methodologies, ready for the lab
Contains vital implementation advice from experts in the field
Captures the current state of this ever-developing and powerful technology
Features practical methodologies, ready for the lab
Contains vital implementation advice from experts in the field
Includes supplementary material: sn.pub/extras
The numerous vital applications of complementary DNA (cDNA) technology have changed dramatically as the technology has advanced over recent years. In cDNA Libraries: Methods and Protocols, expert researchers provide current techniques that reflect the latest advances in the construction and application of cDNA libraries. The first half of the volume covers improved approaches to some of the most basic elements of creating cDNA libraries, while the second half casts a much wider net and includes visionary applications of cDNA technology which were either unforeseen or technically impractical until recently. 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 cutting-edge,
cDNA Libraries: Methods and Protocols serves as an ideal guide to all scientists seeking to advance this important technology and provide answers to the enduring fundamental questions of biology.
Part I: Enhanced Approaches to Inherent Problems
1. Isolation of Fungal Infection Structures from Plant Tissue by Flow Cytometry for Cell-Specific Tranillegalscriptome Analysis
Hiroyuki Takahara, Elmar Endl, and Richard O´Connell
2. Preparation of a High Quality cDNA Library from a Single-Cell Quantity of mRNA Using Chum-RNA
Hiroshi Nojima and Takahiro Tougan
3. Construction of a Full-Length cDNA Library from Castor Endosperm for High-throughput Functional Screening
Chaofu Lu, James G. Wallis, and John Browse
4. Full-Length Tranillegalscriptome Analysis Using a Bias-Free cDNA Library Prepared with the Vector-Capping Method
Seishi Kato, Mio Oshikawa, and Kuniyo Ohtoko
5. Construction of Improved Yeast Two-Hybrid Libraries
Richard H. Maier, Christina J. Maier, and Kamil Önder
6. Normalization of Full-Length Enriched cDNA
Ekaterina A. Bogdanova, Ekaterina V. Barsova, Irina A. Shagina, Alexander Scheglov, Veronika Anisimova, Laura L. Vagner, Sergey A. Lukyanov, and Dmitry A. Shagin
7. Bioinformatic Methods for Finding Differentially Expressed Genes in cDNA Libraries, Applied to the Identification of Tumour Vascular Targets
John M.J. Herbert, Dov J. Stekel, Manuela Mura, Michail Sychev, and Roy Bicknell
Part II: Visionary Applications
8. Enzymatic Production of RNAi Libraries from cDNAs and High-Throughput Selection of Effective shRNA Expression Constructs
Kohtaroh Sugao and Kenzo Hirose
9. Construction of Small RNA cDNA Libraries for High-Throughput Sequencing
Cheng Lu and Vikas Shedge
10. Focusing Mutations Within Random Libraries to Distinct Areas: Protein Domain Library Generation by Overlap Extension (PDLGO)
Andreas Gratz and Joachim Jose
11. Generation of Families of Construct Variants Using Golden Gate Shuffling
Carola Engler and Sylvestre Marillonnet
12. Application of Full-Length cDNA Resources to Gain-of-Function Technology for Characterization of Plant Gene Function
Youichi Kondou, Mieko Higuchi, Takanari Ichikawa, and Minami Matsui
13. Construction of Yeast Surface-Displayed cDNA Libraries
Scott Bidlingmaier and Bin Liu
14. Identification of Protein/Target Molecule Interactions Using Yeast Surface-Displayed cDNA Libraries
Scott Bidlingmaier and Bin Liu
15. SNP Discovery by Tranillegalscriptome Pyrosequencing
W. Brad Barbazuk and Patrick S. Schnable
16. RNA-seq Analysis of Gene Expression and Alternative Splicing by Double-Random Priming Strategy
Michael T. Lovci, Hai-Ri Li, Xiang-dong Fu, and Gene W. Yeo
17. Generation of a Large Catalog of Unique Tranillegalscripts for Whole-Genome Expression Analysis in Non-Model Species
Diana Bellin, Alberto Ferrarini, and Massimo Delledonne
The numerous vital applications of complementary DNA (cDNA) technology have changed dramatically. In this book, expert researchers provide current techniques that reflect the latest advances in the construction and application of cDNA libraries.
The numerous vital applications of complementary DNA (cDNA) technology have changed dramatically as the technology has advanced over recent years. In cDNA Libraries: Methods and Protocols, expert researchers provide current techniques that reflect the latest advances in the construction and application of cDNA libraries. The first half of the volume covers improved approaches to some of the most basic elements of creating cDNA libraries, while the second half casts a much wider net and includes visionary applications of cDNA technology which were either unforeseen or technically impractical until recently. Written in the highly successful Methods in Molecular Biology(TM) 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 cutting-edge,
cDNA Libraries: Methods and Protocols serves as an ideal guide to all scientists seeking to advance this important technology and provide answers to the enduring fundamental questions of biology.
Isolation of Fungal Infection Structures from Plant Tissue by Flow Cytometry for Cell-Specific Tranillegalscriptome Analysis.- Preparation of a High Quality cDNA Library from a Single-Cell Quantity of mRNA Using Chum-RNA.- Construction of a Full-Length cDNA Library from Castor Endosperm for High-throughput Functional Screening.- Full-Length Tranillegalscriptome Analysis Using a Bias-Free cDNA Library Prepared with the Vector-Capping Method.- Construction of Improved Yeast Two-Hybrid Libraries.- Normalization of Full-Length Enriched cDNA.- Bioinformatic Methods for Finding Differentially Expressed Genes in cDNA Libraries, Applied to the Identification of Tumour Vascular Targets.- Enzymatic Production of RNAi Libraries from cDNAs and High-Throughput Selection of Effective shRNA Expression Constructs.- Construction of Small RNA cDNA Libraries for High-Throughput Sequencing.- Focusing Mutations Within Random Libraries to Distinct Areas: Protein Domain Library Generation by Overlap Extension (PDLGO).- Generation of Families of Construct Variants Using Golden Gate Shuffling.- Application of Full-Length cDNA Resources to Gain-of-Function Technology for Characterization of Plant Gene Function.- Construction of Yeast Surface-Displayed cDNA Libraries.- Identification of Protein/Target Molecule Interactions Using Yeast Surface-Displayed cDNA Libraries.- SNP Discovery by Tranillegalscriptome Pyrosequencing.- RNA-seq Analysis of Gene Expression and Alternative Splicing by Double-Random Priming Strategy.- Generation of a Large Catalog of Unique Tranillegalscripts for Whole-Genome Expression Analysis in Non-Model Species.
Inhaltsverzeichnis
Part I: Enhanced Approaches to Inherent Problems
1. Isolation of Fungal Infection Structures from Plant Tissue by Flow Cytometry for Cell-Specific Tranillegalscriptome Analysis
Hiroyuki Takahara, Elmar Endl, and Richard O¿Connell
2. Preparation of a High Quality cDNA Library from a Single-Cell Quantity of mRNA Using Chum-RNA
Hiroshi Nojima and Takahiro Tougan
3. Construction of a Full-Length cDNA Library from Castor Endosperm for High-throughput Functional Screening
Chaofu Lu, James G. Wallis, and John Browse
4. Full-Length Tranillegalscriptome Analysis Using a Bias-Free cDNA Library Prepared with the Vector-Capping Method
Seishi Kato, Mio Oshikawa, and Kuniyo Ohtoko
5. Construction of Improved Yeast Two-Hybrid Libraries
Richard H. Maier, Christina J. Maier, and Kamil Önder
6. Normalization of Full-Length Enriched cDNA
Ekaterina A. Bogdanova, Ekaterina V. Barsova, Irina A. Shagina, Alexander Scheglov, Veronika Anisimova, Laura L. Vagner, Sergey A. Lukyanov, and Dmitry A. Shagin
7. Bioinformatic Methods for Finding Differentially Expressed Genes in cDNA Libraries, Applied to the Identification of Tumour Vascular Targets
John M.J. Herbert, Dov J. Stekel, Manuela Mura, Michail Sychev, and Roy Bicknell
Part II: Visionary Applications
8. Enzymatic Production of RNAi Libraries from cDNAs and High-Throughput Selection of Effective shRNA Expression Constructs
Kohtaroh Sugao and Kenzo Hirose
9. Construction of Small RNA cDNA Libraries for High-Throughput Sequencing
Cheng Lu and Vikas Shedge
10. Focusing Mutations Within Random Libraries to Distinct Areas: Protein Domain Library Generation by Overlap Extension (PDLGO)
Andreas Gratz and Joachim Jose
11. Generation of Families of Construct Variants Using Golden Gate Shuffling
Carola Engler and Sylvestre Marillonnet
12. Application of Full-Length cDNA Resources to Gain-of-Function Technology for Characterization of Plant Gene Function
Youichi Kondou, Mieko Higuchi, Takanari Ichikawa, and Minami Matsui
13. Construction of Yeast Surface-Displayed cDNA Libraries
Scott Bidlingmaier and Bin Liu
14. Identification of Protein/Target Molecule Interactions Using Yeast Surface-Displayed cDNA Libraries
Scott Bidlingmaier and Bin Liu
15. SNP Discovery by Tranillegalscriptome Pyrosequencing
W. Brad Barbazuk and Patrick S. Schnable
16. RNA-seq Analysis of Gene Expression and Alternative Splicing by Double-Random Priming Strategy
Michael T. Lovci, Hai-Ri Li, Xiang-dong Fu, and Gene W. Yeo
17. Generation of a Large Catalog of Unique Tranillegalscripts for Whole-Genome Expression Analysis in Non-Model Species
Diana Bellin, Alberto Ferrarini, and Massimo Delledonne
Klappentext
The numerous vital applications of complementary DNA (cDNA) technology have changed dramatically as the technology has advanced over recent years. In cDNA Libraries: Methods and Protocols, expert researchers provide current techniques that reflect the latest advances in the construction and application of cDNA libraries. The first half of the volume covers improved approaches to some of the most basic elements of creating cDNA libraries, while the second half casts a much wider net and includes visionary applications of cDNA technology which were either unforeseen or technically impractical until recently. Written in the highly successful Methods in Molecular Biology(TM) 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 cutting-edge, cDNA Libraries: Methods and Protocols serves as an ideal guide to all scientists seeking to advance this important technology and provide answers to the enduring fundamental questions of biology.
Captures the current state of this ever-developing and powerful technology
Features practical methodologies, ready for the lab
Contains vital implementation advice from experts in the field
Includes supplementary material: sn.pub/extras
