PCR Protocols / Edition 3

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Known for flexibility and robustness, PCR techniques continue to improve through numerous developments, including the identification of thermostable DNA polymerases which exhibit a range of properties to suit given applications. PCR Prools, Third Edition selects recently developed tools and tricks, contributed by field-leading authors, for the significant value that they add to more generally established methods. Along with the cutting-edge methodologies, this volume describes many core applications, such as PCR cloning and sequencing, expression, copy number or methylation profile analysis, ‘DNA fingerprinting’, diagnostics, protein engineering, interaction screening as well as a chapter highlighting workflow considerations and contamination control, crucial for all PCR methods. Written in the highly successful Methods in Molecular Biology™ series format, chapters include introductions to their respective topics, lists of the necessary reagents and materials, step-by-step, readily reproducible laboratory prools, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and up-to-date, PCR Prools, Third Edition seeks to further elucidate this essential technique while also providing core principles with broad applications for scientists of all backgrounds.

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Editorial Reviews

Doody's Review Service
Reviewer: Omer Iqbal, MD, FACC, FESC (Loyola University Medical Center)
Description: This book describes recently developed tools such as PCR cloning and sequencing, expression, copy number or methylation profile analysis, DNA fingerprinting, diagnostics, protein engineering interaction screening, and other important techniques crucial for all PCR methods.
Purpose: The purpose is to provide scientists with latest tools necessary for PCR methods.
Audience: Students and researchers interested in learning latest methods in PCR will find this book extremely useful.
Features: The book is divided into five parts. Part I covers PCR husbandry and includes an interesting chapter on the setup of a PCR laboratory. Part II covers cloning and sequencing, part III, expression, epigenetic and copy number analysis, part IV, probes and assays, and part V, engineering and screening. Each chapter uses the same format: an introduction, lists of necessary reagents and materials, step-by-step readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Assessment: This book presents cutting-edge methodologies crucial for all PCR methods. Given the rapid advances in the field of PCR methods, this third edition is essential and justifies replacement of the previous edition.
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Product Details

  • ISBN-13: 9781607619437
  • Publisher: Springer-Verlag New York, LLC
  • Publication date: 10/28/2010
  • Series: Methods in Molecular Biology Series, #687
  • Edition description: 3rd ed. 2011
  • Edition number: 3
  • Pages: 348
  • Product dimensions: 0.81 (w) x 7.00 (h) x 10.00 (d)

Table of Contents

Part I: PCR Husbandry

1. Setup of a PCR Laboratory
Zaheer Khan

Part II: Cloning and Sequencing

2. Long Range PCR with a DNA Polymerase Fusion
Holly H. Hogrefe and Michael C. Borns

3. Isolation of Genomic Insertion Sites of Proviruses Using Splinkerette-PCR-Based Procedures
Bin Yin

4. Lariat-Dependent Nested PCR for Flanking Sequence Determination
Daniel J. Park

5. CODEHOP PCR and CODEHOP PCR Primer Design
Jeannette P. Staheli, Richard Boyce, Dina Kovarik, and Timothy M. Rose

6. Sequencing of Difficult DNA Regions by SAM Sequencing
Keith R. Mitchelson

Part III: Expression, Epigenetic and Copy Number Analysis

7. A Global Single-Cell cDNA Amplification Method for Quantitative Microarray Analysis
Kazuki Kurimoto and Mitinori Saitou

8. Quantitation of MicroRNAs by Real-Time RT-qPCR
Caifu Chen, Ruoying Tan, Linda Wong, Richard Fekete, and Jason Halsey

9. High-Throughput SuperSAGE
Hideo Matsumura, Kentaro Yoshida, Shujun Luo, Detlev H. Krüger, Günter Kahl, Gary P. Schroth, and Ryohei Terauchi

10. Deep Cap Analysis of Gene Expression (Deep CAGE)
Junpei Kurosawa, Hiromi Nishiyori, and Yoshihide Hayashizaki

11. Linking Emulsion PCR Haplotype Analysis
James G. Wetmur and Jia Chen

12. PAP-LMPCR: An Improved, Sequence-Selective Method for the In vivo Analysis of Transcription Factor Occupancy and Chromatin Fine Structure
Richard Ingram, Arthur Riggs, and Constanze Bonifer

13. The Many Faces of MLPA
Thomas Ohnesorg, Erin Turbitt, and Stefan J. White

14. Assessing Gene-Specific Methylation Using HRM-Based Analysis
Ee Ming Wong and Alexander Dobrovic

Part IV: Probes and Assays

15. Alu PCR
Maurizio Cardelli

16. Asynchronous PCR
Caifu Chen, David Ruff, and Jason Halsey

17. Novel Applications of PCR through the Use of DNA Substrates
Stuart M. Wilson

18. Enhanced Solid Phase PCR for Increased Loading of Amplicon onto Solid Support
Daniel J. Park

19. Application of Blocking Oligonucleotides to Improve Signal to Noise Ratio in a PCR
Hege Vestheim, Bruce E. Deagle, and Simon N. Jarman

Part V: Engineering and Screening

20. Asymmetric Overlap Extension PCR Method for Site-Directed Mutagenesis
Yue-Hua Xiao and Yan Pei

21. Ribosome Display: A Technology for Selecting and Evolving Proteins from Large Libraries
Birgit Dreier and Andreas Plückthun

22. GLOBE: Analysis of DNA-Protein Interaction Analysis
Takaaki Kojima and Hideo Nakano

23. PCR DNA-Array Profiling of DNA-Binding Transcription Factor Activities in Adult Mouse Tissues
Yimin Sun, Jing Cheng, and Keith R. Mitchelson

24. Nucleotide Exchange and Excision Technology (NExT) DNA Shuffling and Directed Evolution
Janina Speck, Sabine C. Stebel, Katja M. Arndt, and Kristian M. Müller

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