Temperature modulation of DHPLC analysis for detection of coexisting constitutional and mosaic sequence variants in TSC2

Emmanuel S. Antonarakis, Julian R. Sampson, Jeremy P. Cheadle

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

Somatic mosaicism is a frequent phenomenon in Mendelian disorders that exhibit a high proportion of new mutations. However, mutant alleles present at low frequency may escape detection. We have previously shown that denaturing high-performance liquid chromatography (DHPLC) at the recommended melt temperature can detect TSC1 and TSC2 mutations in tuberous sclerosis patients with low-level somatic mosaicism, even when direct sequencing cannot identify the causative lesion. Here, we report the use of temperature modulation in DHPLC analysis to facilitate the robust detection of a mosaic mutation, N1643K, in the presence of a coexisting constitutional polymorphism.

Original languageEnglish (US)
Pages (from-to)161-164
Number of pages4
JournalJournal of Biochemical and Biophysical Methods
Volume51
Issue number2
DOIs
StatePublished - Apr 18 2002
Externally publishedYes

Bibliographical note

Funding Information:
We wish to thank Julie Maynard, Nada Al-Tassan, Peter Davies, and Nick Fleming for technical assistance. This work was supported by grants from the Tuberous Sclerosis Association (UK) and the Tuberous Sclerosis Alliance. E.S.A. was partly funded by the Wolfson Intercalated Award Programme (Royal College of Physicians).

Keywords

  • DHPLC
  • Mosaicism
  • TSC2
  • Temperature modulation

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