Generation of cDNA Libraries Methods and Protocols

About this book

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.

Contents

Complementary DNA Libraries: An Overview Shao-Yao Ying Rapid Amplification of cDNA Ends (RACE) 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 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 Agrose 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 Zhan, Qiu-Hua Huang, and Ze-Guang Han Screening Poly(dA/dT) cDNAs for Gene Identification San Ming Wang, Scott C. Fears, Lin Zhang, Jian-Jun Chen, and Janet D. Rowley Generation of Longer cDNA Fragments from SAGE Tags for Gene Identification Jianjun 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 Rebecca L, Mullinax, Bonnie Wu, and Joseph A. Sorge Future Perspectives Shao-Yao Ying

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