Generation of Cdna Libraries

Generation of Cdna Libraries : Methods and Protocols

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Since its invention and subsequent development nearly 20 years ago, po- merase chain reaction (PCR) has been extensively utilized to identify numerous gene probes in vitro and in vivo. However, attempts to generate complete and full-length complementary cDNA libraries were, for the most part, fruitless and remained elusive until the last decade, when simple and rapid methods were developed. With current decoding and potential application of human genome information to genechips, there are urgent needs for identification of functional significance of these decoded gene sequences. Inherent in bringing these app- cations to fruition is the need to generate a complete and full-length cDNA library for potential functional assays of specific gene sequences. Generation of cDNA Libraries: Methods and Protocols serves as a laboratory manual on the evolution of generation of cDNA libraries, covering both ba- ground information and step-by-step practical laboratory recipes for which p- tocols, reagents, operational tips, instrumentation, and other requirements are detailed. The first chapter of the book is an overview of the basics of generating cDNA libraries, which include the following: (a) the definition of a cDNA library, (b) different kinds of cDNA libraries, (c) differences between methods for cDNA library generation using conventional approaches and novel stra- gies, including reverse generation of RNA repertoires from cDNA libraries, and (d) the quality of cDNA more

Product details

  • Hardback | 338 pages
  • 154.9 x 231.1 x 25.4mm | 557.93g
  • Humana Press Inc.
  • Totowa, NJ, United States
  • English
  • 2003 ed.
  • biography
  • 1588290662
  • 9781588290663

Review quote

...the ready-for-use protocols are excellent...A good buy. - Cell Biochem Functshow more

Back cover copy

With the decoding of the human genome and the potential application of its information to gene chips, the ability to generate cDNA libraries for functional assays of specific gene sequences has become critical in gene manipulation and genetic research. In Generation of cDNA Libraries: Methods and Protocols, expert researchers and inventors in the field describe their own proven techniques for generating cDNA/mRNA libraries to identify the functions of specific decoded gene sequences. A wide variety of techniques is presented for enhancing the generation of complete and full-length libraries, and for confirming the quality of the cDNAs generated. Among the applications detailed are electrophoresis, Northern blotting, single-cell microarray analysis, subtractive hybridization, subtractive cloning, gene cloning, and peptide library generation. Each method includes background information, step-by-step instructions, a list of reagents, operational tips, and notes on instrumentation. A background tutorial provides essential information: the definition of a cDNA library, the various types of cDNA libraries, the differences between methods for cDNA library generation using either conventional approaches or novel strategies, and the quality of cDNA libraries. State-of-the-art and highly practical, Generation of cDNA Libraries: Methods and Protocols provides a variety of powerful and readily reproducible techniques for the generation of the entire range of complete, full-length cDNA/mRNA libraries needed in today's forefront genetic more

Table of contents

Complementary DNA Libraries: An Overview Shao-Yao Ying Rapid Amplification of cDNA Ends Yue Zhang cDNA Generation on Paramagnetic Beads Zhaohui Wang and Michael G. K. Jones Construction of a Normalized cDNA Library by mRNA-cDNA Hybridization and Subtraction Ye-Guang Chen Amplification of cDNA Ends Using PCR Suppression Effect and Step-Out PCR Mikhail V. Matz, Naila O. Alieva, Alex Chenchik, and Sergey Lukyanov Use of Inverse PCR to Clone cDNA Ends Sheng-He Huang, Steven H. M. Chen, and Ambrose Y. Jong Construction of Size-Fractionated cDNA Library Assisted by an In Vitro Recombination Reaction Osamu Ohara Construction of a Full-Length Enriched and 5'-End Enriched cDNA Library Using the Oligo-Capping Method Yutaka Suzuki and Sumio Sugano cDNA Library Construction Using In Vitro Transcriptional Amplification Shi-Lung Lin and Henry Ji Amplification of Representative cDNA Pools from Microscopic Amounts of Animal Tissue Mikhail V. Matz Single-Cell cDNA Library Construction Using Cycling aRNA Amplification Shi-Lung Lin mRNA/cDNA Library Construction Using RNA-Polymerase Cycling Reaction Shi-Lung Lin and Shao-Yao Ying Quality Assessment of cDNA Libraries Hans-Jurgen Fulle Assessment of the Quality of mRNA Libraries by Agarose Gel Electrophoresis Tsen-Yin Lin and Shao-Yao Ying PACS RT-PCR: A Method for the Generation and Measurement of any Poly(A)-Containing mRNA Not Affected by Contaminating Genomic DNA Igor Nepluev and Rodney J. Folz Single-Cell mRNA Library Analysis by Northern Blot Hybridization Shi-Lung Lin Generation of cDNA Libraries for Profiling Gene Expression of Given Tissues or Cells Xin Zhang, Qiu-Hua Huang, and Ze-Guang Han Screening Poly [dA/dT(-)] cDNA for Gene Identification San MingWang, Scott C. Fears, Lin Zhang, Jian-Jun Chen, and Janet D. Rowley Generation of Longer cDNA Fragments from SAGE Tags for Gene Identification Jian-Jun Chen, Sanggyu Lee, Guolin Zhou, Janet D. Rowley, and San Ming Wang Generation of Full-Length cDNA Libraries Enriched for Differentially Expressed Genes Bakhyt Zhumabayeva, Cynthia Chang, Joseph McKinley, Luda Diatchenko, and Paul D. Siebert Subtractive Hybridization for the Identification of Differentially Expressed Genes Using Uracil-DNA Glycosylase and Mung-Bean Nuclease Tsen-Yin Lin and Shao-Yao Ying Subtractive Cloning of Differential Genes Using RNA-PCR Shao-Yao Ying and Shi-Lung Lin Strategy for Construction of a cDNA Encoding a Repetitive Amino Acid Sequence Masahiro Asada and Toru Imamura Preparing Lambda Libraries for Expression of Proteins in Prokaryotes or Eukaryotes Rebecca L. Mullinax and Joseph A. Sorge Peptide Library Construction from RNA-PCR-Derived RNAs Shi-Lung Lin Identifying Interacting Proteins in an Escherichia coli-Based Two-Hybrid System Bonnie Wu, Rebecca L. Mullinax, and Joseph A. Sorge Future Perspectives, Shao-Yao Ying Indexshow more