Generation of cDNA libraries : methods and protocols / edited by Shao-Yao Ying.Material type: TextSeries: Methods in molecular biology (Clifton, N.J.) ; v. 221.Publication details: Totowa, N.J. : Humana Press, ©2003. Description: 1 online resource (xiii, 335 pages) : illustrationsContent type: text Media type: computer Carrier type: online resourceISBN: 9781592593590; 1588290662; 9781588290663; 1592593593; 1280842393; 9781280842399; 9786610842391; 6610842396; 9781591593591; 159159359XSubject(s): Antisense DNA -- Laboratory manuals | DNA, Antisense | Combinatorial Chemistry Techniques -- methods | Gene Library | MEDICAL -- Genetics | Antisense DNA | DNA | Kloneren | Genetische manipulatie | Wetenschappelijke techniekenGenre/Form: Laboratory Manual. | Electronic books. | Laboratory manuals. Additional physical formats: Print version:: Generation of cDNA libraries.DDC classification: 611/.01816 LOC classification: QP624.5.A57 | G46 2003NLM classification: W1 | QU 25Online resources: Click here to access online
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Includes bibliographical references and index.
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 [and others] -- 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-Jürgen Fülle -- 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 Ming Wang [and others] -- Generation of longer cDNA fragments from SAGE tags for gene identification / Jian-Jun Chen [and others] -- Generation of full-length cDNA libraries enriched for differentiatlly expressed genes / Bakhyt Zhumabayeva [and others] -- 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.
Print version record.
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 research.