Generation of cDNA libraries : methods and protocols / edited by Shao-Yao Ying.

Format:

Book

Series:

Methods in molecular biology (Clifton, N.J.) ; 221.

Methods in molecular biology ; 221

Language:

English

Subjects (All):

Antisense DNA--Laboratory manuals.

Antisense DNA.

DNA, Antisense.

Medical Subjects:

DNA, Antisense.

Genre:

Laboratory manuals.

Physical Description:

xiii, 335 pages : illustrations ; 24 cm.

Place of Publication:

Totowa, NJ : Humana Press, 2003.

Summary:

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.

Contents:

1 Complementary DNA Libraries: An Overview / Shao-Yao Ying 1

2 Rapid Amplification of cDNA Ends / Yue Zhang 13

3 cDNA Generation on Paramagnetic Beads / Zhaohui Wang, Michael G. K. Jones 25

4 Construction of a Normalized cDNA Library by mRNA-cDNA Hybridization and Subtraction / Ye-Guang Chen 33

5 Amplification of cDNA Ends Using PCR Suppression Effect and Step-Out PCR / Mikhail V. Matz, Naila O. Alieva, Alex Chenchik, Sergey Lukyanov 41

6 Use of Inverse PCR to Clone cDNA Ends / Sheng-He Huang, Steven H. M. Chen, Ambrose Y. Jong 51

7 Construction of Size-Fractionated cDNA Library Assisted by an In Vitro Recombination Reaction / Osamu Ohara 59

8 Construction of a Full-Length Enriched and 5'-End Enriched cDNA Library Using the Oligo-Capping Method / Yutaka Suzuki, Sumio Sugano 73

9 cDNA Library Construction Using In Vitro Transcriptional Amplification / Shi-Lung Lin, Henry Ji 93

10 Amplification of Representative cDNA Pools from Microscopic Amounts of Animal Tissue / Mikhail V. Matz 103

11 Single-Cell cDNA Library Construction Using Cycling aRNA Amplification / Shi-Lung Lin 117

12 mRNA/cDNA Library Construction Using RNA-Polymerase Cycling Reaction / Shi-Lung Lin, Shao-Yao Ying 129

13 Quality Assessment of cDNA Libraries / Hans-Jurgen Fulle 145

14 Assessment of the Quality of mRNA Libraries by Agarose Gel Electrophoresis / Tsen-Yin Lin, Shao-Yao Ying 155

15 PACS RT-PCR: A Method for the Generation and Measurement of any Poly(A)-Containing mRNA Not Affected by Contaminating Genomic DNA / Igor Nepluev, Rodney J. Folz 161

16 Single-Cell mRNA Library Analysis by Northern Blot Hybridization / Shi-Lung Lin 169

17 Generation of cDNA Libraries for Profiling Gene Expression of Given Tissues or Cells / Xin Zhang, Qiu-Hua Huang, Ze-Guang Han 179

18 Screening Poly [dA/dT(-)] cDNA for Gene Identification / San Ming Wang, Scott C. Fears, Lin Zhang, Jian-Jun Chen, Janet D. Rowley 197

19 Generation of Longer cDNA Fragments from SAGE Tags for Gene Identification / Jian-Jun Chen, Sanggyu Lee, Guolin Zhou, Janet D. Rowley, San Ming Wang 207

20 Generation of Full-Length cDNA Libraries Enriched for Differentially Expressed Genes / Bakhyt Zhumabayeva, Cynthia Chang, Joseph McKinley, Luda Diatchenko, Paul D. Siebert 223

21 Subtractive Hybridization for the Identification of Differentially Expressed Genes Using Uracil-DNA Glycosylase and Mung-Bean Nuclease / Tsen-Yin Lin, Shao-Yao Ying 239

22 Subtractive Cloning of Differential Genes Using RNA-PCR / Shao-Yao Ying, Shi-Lung Lin 253

23 Strategy for Construction of a cDNA Encoding a Repetitive Amino Acid Sequence / Masahiro Asada, Toru Imamura 261

24 Preparing Lambda Libraries for Expression of Proteins in Prokaryotes or Eukaryotes / Rebecca L. Mullinax, Joseph A. Sorge 271

25 Peptide Library Construction from RNA-PCR-Derived RNAs / Shi-Lung Lin 289

26 Identifying Interacting Proteins in an Escherichia coli-Based Two-Hybrid System / Bonnie Wu, Rebecca L. Mullinax, Joseph A. Sorge 295

27 Future Perspectives / Shao-Yao Ying 311.

Notes:

Includes bibliographical references and index.

ISBN:

1588290662

OCLC:

51178265