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
UR - http://www.scopus.com/inward/record.url?scp=0037129175&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0037129175&partnerID=8YFLogxK
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 -