Lehmann U., Tost J. (Eds.) Pyrosequencing. Methods and Protocols

Humana Press, 2015 — 2nd Ed. — 414p. — (Methods in Molecular Biology 1315) — ISBN: 978-1-4939-2715-9 (eBook), 978-1-4939-2714-2 (Hardcover).The primary purpose of this volume is to demonstrate the range of applications of the Pyrosequencing technology in research and diagnostics and to provide detailed protocols. Beginning with an up-to-date overview of the biochemistry, the volume continues with quantitative analysis of genetic variation, ratio of expressed alleles at the RNA level, analysis of DNA methylation, global DNA methylation assays, specialized applications for DNA methylation analysis including loss of imprinting, single blastocyst analysis, allele-specific DNA methylation patterns, DNA methylation patterns associated with specific histone modifications. The volume further details tools and protocols for the detection of viruses and bacteria, and genetic and epigenetic analyses for forensics using Pyrosequencing. As a volume in the highly successful Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols and tips on troubleshooting and avoiding known pitfalls.
Comprehensive and adaptable, Pyrosequencing: Methods and Protocols, Second Edition will greatly aid doctorial students, postdoctoral investigators and research scientists studying different aspects of genetics and cellular and molecular biology.The History of Pyrosequencing
PyroMark Instruments, Chemistry, and Software for Pyrosequencing Analysis
Software-Based Pyrogram Evaluation
Quantitative Validation and Quality Control of Pyrosequencing Assays
Extended KRAS and NRAS Mutation Profiling by Pyrosequencing
Universal BRAF State Detection by the Pyrosequencing-Based U-BRAF^V600 Assay
Pyrosequencing-Based Identification of Low-Frequency Mutations Enriched Through Enhanced-ice-COLD-PCR
Analysis of Mutational Hotspots in Routinely Processed Bone Marrow Trephines by Pyrosequencing
Analysis of Copy Number Variation by Pyrosequencing Using Paralogous Sequences
Prenatal Diagnosis of Chromosomal Aneuploidies by Quantitative Pyrosequencing
HLA-B and HLA-C Supratyping by Pyrosequencing
Allele Quantification Pyrosequencing at Designated SNP Sites to Detect Allelic Expression Imbalance and Loss-of-Heterozygosity
Quantitative DNA Methylation Analysis by Pyrosequencing
Quantitative Methylation Analysis of the PCDHB Gene Cluster
Assessment of Changes in Global DNA Methylation Levels by Pyrosequencing of Repetitive Elements
Global Analysis of DNA 5-Methylcytosine Using the Luminometric Methylation Assay, LUMA
Limiting Dilution Bisulfite Pyrosequencing: A Method for Methylation Analysis of Individual DNA Molecules in a Single or a Few Cells
Detection of Loss of Imprinting by Pyrosequencing
Analysis of DNA Methylation Patterns in Single Blastocysts by Pyrosequencing
Allele-Specific DNA Methylation Detection by Pyrosequencing