Advances and Clinical Practice in Pyrosequencing [Paperback]

The aim of this book is to improve pyrosequencing protocols as well as instrumentation for better clinical use by describing improvements and novel applications of pyrosequencing technology.  Divided into five parts, the books thirty chapters explore advances in pyrosequencing template preparation, pyrosequencing technology innovations, multiplex pyrosequencing based on barcodes, the miniaturization of pyrosequencing equipment, as well as various applications. As part of the Springer Protocols program, chapters contain the kind of detail and practical implementation advice to guarantee successful results in the lab.

Comprehensive and thorough, Advances and Clinical Practice in Pyrosequencing serves as a valuable reference for researchers who are engaged in personalized medicine, disease control, and DNA diagnosis in numerous other fields.

Preparation of single-stranded DNA for pyrosequencing by LATE-PCR.- A simplified protocol for preparing pyrosequencing templates based on LATE-PCR using whole blood as starting material directly.- Improvement of LATE-PCR to prepare pyrosequencing template.- Pyrosequencing-templates generated by nicking PCR products with nicking endonuclease.- Pyrosequencing-templates generated by asymmetric Nucleic Acid Sequence-Based Amplification (asymmetric-NASBA).- Gel immobilization of acrylamide-modified single-stranded DNA template for pyrosequencing.- Multiplex PCR based on a universal biotinylated primer to generate templates for pyrosequencing.- A novel pyrosequencing principle based on AMP-PPDK reaction for improving the detection limit.- Pyrosequencing chemistry coupled with modified primer extension reactions for quantitative detection of allele frequency in a pooled sample.- A gel-free SNP genotyping method based on pyrosequencing chemistry coupled with modified primer extension reactionsdirectly from double-stranded PCR products.- Improvement of pyrlÓ.