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

doi:10.1016/S0165-022X(02)00011-8

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 journal › Article › peer-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 language	English (US)
Pages (from-to)	161-164
Number of pages	4
Journal	Journal of Biochemical and Biophysical Methods
Volume	51
Issue number	2
DOIs	https://doi.org/10.1016/S0165-022X(02)00011-8
State	Published - Apr 18 2002
Externally published	Yes

Keywords

DHPLC
Mosaicism
TSC2
Temperature modulation

ASJC Scopus subject areas

Biophysics
Biochemistry

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10.1016/S0165-022X(02)00011-8

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title = "Temperature modulation of DHPLC analysis for detection of coexisting constitutional and mosaic sequence variants in TSC2",

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.",

keywords = "DHPLC, Mosaicism, TSC2, Temperature modulation",

author = "Antonarakis, {Emmanuel S.} and Sampson, {Julian R.} and Cheadle, {Jeremy P.}",

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).",

year = "2002",

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doi = "10.1016/S0165-022X(02)00011-8",

language = "English (US)",

volume = "51",

pages = "161--164",

journal = "Journal of Biochemical and Biophysical Methods",

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TY - JOUR

T1 - Temperature modulation of DHPLC analysis for detection of coexisting constitutional and mosaic sequence variants in TSC2

AU - Antonarakis, Emmanuel S.

AU - Sampson, Julian R.

AU - Cheadle, Jeremy P.

N1 - 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).

PY - 2002/4/18

Y1 - 2002/4/18

N2 - 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.

AB - 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.

KW - DHPLC

KW - Mosaicism

KW - TSC2

KW - Temperature modulation

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U2 - 10.1016/S0165-022X(02)00011-8

DO - 10.1016/S0165-022X(02)00011-8

M3 - Article

C2 - 12062115

AN - SCOPUS:0037129175

SN - 0165-022X

VL - 51

SP - 161

EP - 164

JO - Journal of Biochemical and Biophysical Methods

JF - Journal of Biochemical and Biophysical Methods

IS - 2

ER -

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

Abstract

Keywords

ASJC Scopus subject areas

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