Pyrosequencing for the rapid and efficient quantification of allele-specific expression

Bing Yang, Jennifer Wagner, Tianyu Yao, Nathan Damaschke, David F. Jarrard

Research output: Contribution to journalArticlepeer-review

2 Scopus citations

Abstract

We have developed a rapid and sensitive quantitative assay for the measurement of individual allelic ratios. This assay minimizes time and labor, the need for special restriction endonuclease enzymes for polymorphic sites, and avoids heteroduplex formation seen with traditional quantitative PCR-based methods. It has improved sensitivity compared to other methods and is capable of distinguishing 1% differences in allelic expression. This assay, termed Pyrosequencing for Imprinted Expression (PI E), involves the use of an intron-crossing PCR primer to generate the first PCR product. We applied the assay to analyze Insulin-like Growth Factor-2 (IGF2) imprinting in both human and mouse prostate tissues.

Original languageEnglish (US)
Pages (from-to)1039-1042
Number of pages4
JournalEpigenetics
Volume8
Issue number10
DOIs
StatePublished - Oct 2013
Externally publishedYes

Keywords

  • Epigenetic
  • IGF2
  • Imprinting
  • Intron-crossing primer
  • Pyrosequencing

ASJC Scopus subject areas

  • Molecular Biology
  • Cancer Research

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