PCR primer : a laboratory manual / edited by Carl W. Dieffenbach, Gabriela S. Dveksler.

Format:

Book

Contributor:

Dieffenbach, Carl W.

Dveksler, Gabriela S.

Language:

English

Subjects (All):

Polymerase chain reaction--Laboratory manuals.

Polymerase chain reaction.

Polymerase Chain Reaction--methods.

Medical Subjects:

Polymerase Chain Reaction--methods.

Genre:

Laboratory manuals.

Physical Description:

xi, 520 pages : illustrations ; 29 cm

Edition:

Second edition.

Other Title:

Polymerase chain reaction primer

Place of Publication:

Cold Spring Harbor, N.Y. : Cold Spring Harbor Laboratory Press, [2003]

Summary:

The Polymerase Chain Reaction (PCR) technique was invented nearly 20 years ago. Its subsequent variations and applications were many and varied, and today molecular biology, clinical, and forensic laboratories make almost daily use of PCR. This second edition of the much-praised PCR Primer: A Laboratory Manual updates the tried-and-true methods and presents the advances made in the 10 years since the first edition. After introducing the basics for PCR and methods of sample preparation, PCR Primer provides laboratory-tested protocols for RT-PCR methods, detection of PCR products, analysis of differential expression, cloning, and mutagenesis. These step-by-step methods include extensive background information, as well as valuable troubleshooting information provided by the leading experts in this technology. This manual is a comprehensive and reliable source of the full range of PCR methods for novices and experienced investigators alike.

Contents:

1 Setting Up a PCR Laboratory / Theodore E. Mifflin 5

2 Dealing with Carryover Contamination in PCR: An Enzymatic Strategy / Kyusung Park, Jun Lee 15

3 High-Fidelity PCR Enzymes / Holly H. Hogrefe, Michael C. Borns 21

4 Optimization and Troubleshooting in PCR / Kenneth H. Roux 35

5 PCR Amplification of Highly GC-rich Regions / Lise Lotte Hansen, Just Justesen 43

6 Tips for Long and Accurate PCR / Wayne M. Barnes 53

7 PCR Primer Design / Arun Apte, Saurabha Daniel 61

8 Nonradioactive Cycle Sequencing of PCR-amplified DNA / Susan R. Haynes 75

2 Sample Preparation 83

9 DNA Purification / Craig Smith, Paul Otto, Rex Bitner, Gary Shiels 87

10 RNA Purification / Michael A. Connolly, Peter A. Clausen, James G. Lazar 117

11 In Situ Localization of Gene Expression Using Laser Capture Microdissection / Motoko Morimoto, Masahiro Morimoto, Jeannette Whitmire, Robert A. Star, Joseph F. Urban, Jr., William C. Gause 135

12 Strategies for Overcoming PCR Inhibition / Peter Radstrom, Charlotta Lofstrom, Maria Lovenklev, Rickard Knutsson, Petra Wolffs 149

3 RT-PCR Methods 163

13 The Use of Internal Controls for Relative and Competitive Quantitative RT-PCR / Renee M. Horner 167

14 Comparison of Four Real-time PCR Detection Systems: Bio-Rad I-Cycler, ABI 7700, Roche LightCycler, and the Cepheid Smartcycler / Katherine E. Templeton, Eric C.J. Claas 187

15 New Technology for Quantitative PCR / Wolfgang Kusser, Sandrine Javorschi, Martin A. Gleeson 199

16 Amplification of RNA: High-temperature Reverse Transcription and DNA Amplification with a Magnesium-activated Thermostable DNA Polymerase / Edward S. Smith, Alvin K. Li, Alice M. Wang, David H. Gelfand, Thomas W. Myers 211

4 Detection of PCR Products: Specialized Applications 221

17 Loss of Heterozygosity, a Multiplex PCR Method to Define Narrow Deleted Chromosomal Regions of a Tumor Genome / Lise Lotte Hansen, Just Justesen 223

18 Single-strand Conformation Polymorphism Analysis / Anne E. Jedlicka, Steven R. Kleeberger 237

19 Reverse Transcriptase In Situ PCR / Gerard J. Nuovo 251

20 Sensitive and Fast Mutation Detection by Solid-phase Chemical Cleavage of Mismatches / Lise Lotte Hansen, Just Justesen, Andreana Lambrinakos, Richard G.H. Cotton 265

5 Analysis of Differential Gene Expression by PCR 279

21 Applying PCR for Microarray-based Gene Expression Analysis / Joan M. Zakel 281

22 Identification of Differential Genes by Suppression Subtractive Hybridization / Denis V. Rebrikov 297

23 Fluorescent mRNA Differential Display in Gene Expression Analysis / Jeffrey S. Fisher, Jonathan Meade, Jamie Walden, Zhen Guo, Peng Liang 327

6 Cloning by PCR 349

24 PCR-based Method for Screening Libraries / David I. Israel 351

25 Beyond Classic RACE (Rapid Amplification of cDNA Ends) / Michael A. Frohman 359

26 Synthesis and Analysis of DNA Libraries Using PCR / John L. Joyal, Jinwei Jiang, David I. Israel 391

7 Cloning of PCR Products 403

27 Strategies for Cloning PCR Products / Peter D'Arpa 405

28 Bidirectional and Directional Cloning of PCR Products / Gina L. Costa, Michael P. Weiner 421

29 Ribocloning: DNA Cloning and Gene Construction Using PCR Primers Terminated with a Ribonucleotide / Wayne M. Barnes 441

8 Mutagenesis by PCR 451

30 Mutagenic PCR / R. Craig Cadwell, Gerald F. Joyce 453

31 Rapid PCR Site-directed Mutagenesis / Michael P. Weiner, Gina L. Costa 459

32 Mutagenesis and Synthesis of Novel Recombinant Genes Using PCR / Abbe N. Vallejo, Robert J. Pogulis, Larry R. Pease 467

33 Streamlined Gene Assembly PCR / Wayne M. Barnes, Elaine R. Frawley 475

1 Licensing 483

2 Modification of Oligonucleotides for Use in PCR 485.

Notes:

Includes bibliographical references and index.

ISBN:

0879696532

0879696540

OCLC:

52288789