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Main description:
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 Protocols, 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 (TM) series format, chapters include introductions to their respective topics, lists of the necessary reagents and materials, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and up-to-date, PCR Protocols, Third Edition seeks to further elucidate this essential technique while also providing core principles with broad applications for scientists of all backgrounds.
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. Kruger, Gunter 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 Pluckthun
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. Muller
PRODUCT DETAILS
Publisher: Springer (Humana Press Inc.)
Publication date: October, 2010
Pages: 360
Weight: 1860g
Availability: Available
Subcategories: Anaesthetics and Pain, Biochemistry, General Issues, Genetics
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