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Pyrosequencing

Methods and Protocols

  • Book
  • © 2015

Overview

Editors:
  1. Ulrich Lehmann

    1. Medizinische Hochschule Hannover, Institute of Pathology, Hannover, Germany

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  2. Jörg Tost

    1. Laboratory for Epigenetics and Environment, Centre National de Génotypage, CEA- Institut de Génomique, Evry, France

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  • Includes cutting-edge methods and protocols
  • Provides step-by-step detail essential for reproducible results
  • Contains key notes and implementation advice from the experts

Part of the book series: Methods in Molecular Biology (MIMB, volume 1315)

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Table of contents (27 protocols)

  1. Front Matter

    Pages i-xiv
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  2. Introduction

    1. Front Matter

      Pages 1-1
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    2. The History of Pyrosequencing®

      • Pål Nyrén
      Pages 3-15

    3. PyroMark® Instruments, Chemistry, and Software for Pyrosequencing® Analysis

      • Martin Kreutz, Gerald Schock, Julia Kaiser, Norbert Hochstein, Ralf Peist
      Pages 17-27

    4. Software-Based Pyrogram® Evaluation

      • Guoli Chen, Matthew T. Olson, James R. Eshleman
      Pages 29-38

    5. Quantitative Validation and Quality Control of Pyrosequencing® Assays

      • Ulrich Lehmann
      Pages 39-46

  3. Analysis of Genetic Variation

    1. Front Matter

      Pages 47-47
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    2. Extended KRAS and NRAS Mutation Profiling by Pyrosequencing®

      • Andreas Jung
      Pages 49-61

    3. Universal BRAF State Detection by the Pyrosequencing®-Based U-BRAFV600 Assay

      • Alexander Skorokhod
      Pages 63-82

    4. Pyrosequencing®-Based Identification of Low-Frequency Mutations Enriched Through Enhanced-ice-COLD-PCR

      • Alexandre How-Kit, Jörg Tost
      Pages 83-101

    5. Analysis of Mutational Hotspots in Routinely Processed Bone Marrow Trephines by Pyrosequencing®

      • Stephan Bartels, Ulrich Lehmann
      Pages 103-114

    6. Analysis of Copy Number Variation by Pyrosequencing® Using Paralogous Sequences

      • Marianne Kristiansen Kringen
      Pages 115-121

    7. Prenatal Diagnosis of Chromosomal Aneuploidies by Quantitative Pyrosequencing®

      • Hui Ye, Haiping Wu, Yunlong Liu, Bingjie Zou, Tomoharu Kajiyama, Hideki Kambara et al.
      Pages 123-132

    8. HLA-B and HLA-C Supratyping by Pyrosequencing®

      • Irene Vanni, Elisabetta Ugolotti, Patrizia Larghero, Roberto Biassoni
      Pages 133-151

    9. Allele Quantification Pyrosequencing® at Designated SNP Sites to Detect Allelic Expression Imbalance and Loss-of-Heterozygosity

      • Chau-To Kwok, Megan P. Hitchins
      Pages 153-171

  4. Analysis of DNA Methylation Patterns

    1. Front Matter

      Pages 173-173
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    2. Quantitative DNA Methylation Analysis by Pyrosequencing®

      • Jessica Roessler, Ulrich Lehmann
      Pages 175-188

    3. Quantitative Methylation Analysis of the PCDHB Gene Cluster

      • Barbara Banelli, Massimo Romani
      Pages 189-200

    4. Assessment of Changes in Global DNA Methylation Levels by Pyrosequencing® of Repetitive Elements

      • Ali M. Tabish, Andrea A. Baccarelli, Lode Godderis, Timothy M. Barrow, Peter Hoet, Hyang-Min Byun
      Pages 201-207

    5. Global Analysis of DNA 5-Methylcytosine Using the Luminometric Methylation Assay, LUMA

      • Karin Luttropp, Louise K. Sjöholm, Tomas J. Ekström
      Pages 209-219
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Keywords

About this book

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.

Editors and Affiliations

  • Medizinische Hochschule Hannover, Institute of Pathology, Hannover, Germany

    Ulrich Lehmann

  • Laboratory for Epigenetics and Environment, Centre National de Génotypage, CEA- Institut de Génomique, Evry, France

    Jörg Tost

Bibliographic Information

  • Book Title: Pyrosequencing

  • Book Subtitle: Methods and Protocols

  • Editors: Ulrich Lehmann, Jörg Tost

  • Series Title: Methods in Molecular Biology

  • DOI: https://doi.org/10.1007/978-1-4939-2715-9

  • Publisher: Humana New York, NY

  • eBook Packages: Springer Protocols

  • Copyright Information: Springer Science+Business Media New York 2015

  • Hardcover ISBN: 978-1-4939-2714-2Published: 24 June 2015

  • Softcover ISBN: 978-1-4939-4909-0Published: 09 October 2016

  • eBook ISBN: 978-1-4939-2715-9Published: 23 June 2015

  • Series ISSN: 1064-3745

  • Series E-ISSN: 1940-6029

  • Edition Number: 2

  • Number of Pages: XIV, 414

  • Number of Illustrations: 26 b/w illustrations, 57 illustrations in colour

  • Topics: Gene Function

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