Pyrosequencing pp 115-121 | Cite as

Analysis of Copy Number Variation by Pyrosequencing® Using Paralogous Sequences

  • Marianne Kristiansen Kringen
Part of the Methods in Molecular Biology book series (MIMB, volume 1315)


The determination of gene copy numbers of highly similar genes is difficult with conventional PCR-based methods. However, by amplification of similar genes in the same PCR reaction followed by Pyrosequencing®, one may distinguish the genes based on a single-nucleotide difference. The ratio between the peak heights of gene-specific nucleotides obtained in the Pyrosequencing reaction may thereby be used to calculate the relative copy numbers of target genes. This method is easy and cost effective compared to other methods, and allows for the determination of copy numbers of genes that were previously difficult to achieve.

Key words

Copy number variation PCR Pyrosequencing® Paralogous sequences ABCC6 


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.Department of PharmacologyOslo University Hospital, UllevålOsloNorway

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