PCR Protocols

In its 3rd Edition, this book selects recently developed tools and tricks, for the value they add to established methods. The text describes core applications: PCR cloning and sequencing, expression, copy number or methylation profile analysis, and more.

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™ 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.

Presents detailed, independent methods, the core principles of which have broad and varied applications in the lab Describes thoroughly the most recent and exciting fully developed protocols along with tips from the expert contributors Inspires alternative approaches to aid all labs in producing successful results

Inhalt
Setup of a PCR Laboratory.- Long Range PCR with a DNA Polymerase Fusion.- Isolation of Genomic Insertion Sites of Proviruses Using Splinkerette-PCR-Based Procedures.- Lariat-Dependent Nested PCR for Flanking Sequence Determination.- CODEHOP PCR and CODEHOP PCR Primer Design.- Sequencing of Difficult DNA Regions by SAM Sequencing.- A Global Single-Cell cDNA Amplification Method for Quantitative Microarray Analysis.- Quantitation of MicroRNAs by Real-Time RT-qPCR.- High-Throughput SuperSAGE.- Deep Cap Analysis of Gene Expression (Deep CAGE).- Linking Emulsion PCR Haplotype Analysis.- PAP-LMPCR: An Improved, Sequence-Selective Method for the In vivo Analysis of Transcription Factor Occupancy and Chromatin Fine Structure.- The Many Faces of MLPA.- Assessing Gene-Specific Methylation Using HRM-Based Analysis.- Alu PCR.- Asynchronous PCR.- Novel Applications of PCR through the Use of DNA Substrates.- Enhanced Solid Phase PCR for Increased Loading of Amplicon onto Solid Support.- Application of Blocking Oligonucleotides to Improve Signal to Noise Ratio in a PCR.- Asymmetric Overlap Extension PCR Method for Site-Directed Mutagenesis.- Ribosome Display: A Technology for Selecting and Evolving Proteins from Large Libraries.- GLOBE: Analysis of DNA-Protein Interaction Analysis.- PCR DNA-Array Profiling of DNA-Binding Transcription Factor Activities in Adult Mouse Tissues.- Nucleotide Exchange and Excision Technology (NExT) DNA Shuffling and Directed Evolution.