Abstract
The chapter details the methodology for polymerase chain reaction amplification and WAVE denaturing high-performance liquid chromatography (DHPLC) analysis for all coding exons for the gene PTPN11, which is mutated in approx 50% of cases of Noonan Syndrome. Although DNA sequencing is initially required to determine the mutation(s) detected by WAVE (sequencing methods are not described in this chapter), each mutation has its own DHPLC signature, and experienced operatives can determine known mutations on this basis. The new Navigator software has made this process more reliable.
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Sharland, M., Morgan, M., Smith, G., Burch, M., and Patton, M. A. (1993) Genetic counseling in Noonan syndrome. Am. J. Med. Genet. 45, 437–440.
Jamieson, R., van der Burgt, I., Brady, A., et al. (1994) Mapping a gene for Noonan Syndrome to the long arm of chromosome 12. Nat. Genet. 8, 357–360.
Tartaglia, M., Mehler, E. L., Goldberg, R., et al. (2001) Mutations in PTPN11, encoding the protein tyrosine phosphatase SHP-2, cause Noonan syndrome. Nat. Genet. 29, 465–468.
Chen, B., Bronson, R. T., Klaman, L. D., et al. (2000) Mice mutants for Egfr and Shp2 have defective cardiac semilunar valvulogenesis. Nat. Genet. 24, 296–299.
Tartaglia, M., Kalidas, K., Shaw, A., et al. (2002) PTPN11 Mutations in Noonan Syndrome: Molecular Spectrum, Genotype-Phenotype Correlation, and Phenotypic Heterogeneity. Am. J. Hum. Genet. 70, 1555–1563.
Musante, L., Kehl, H. G., Majewski, F., et al. (2003) Spectrum of mutations in PTPN11 and genotype-phenotype correlation in 96 patients with Noonan syndrome and five patients with cardio-facio-cutaneous syndrome. Eur. J. Hum. Genet. 11, 201–206.
Maheshwari, M., Belmont, J., Fernbach, S., et al. (2002) PTPN11 mutations in Noonan syndrome type I: detection of recurrent mutations in exons 3 and 13. Hum. Mutat. 20, 298–304.
Sarkozy, A., Conti, E., Seripa, D., et al. (2203) Correlation between PTPN11 gene mutations and congenital heart defects in Noonan and LEOPARD syndromes. J. Med. Genet. 40, 704–708.
Mogensen, J., Bahl, A., Kubo, T., Elanko, N., Taylor, R., and McKenna, W. J. (2003). Comparison of fluorescent SSCP and denaturing HPLC analysis with direct sequencing for mutation screening in hypertrophic cardiomyopathy. J. Med. Genet. 40, 159.
Jones, A. C., Austin, J., Hansen, N., et al. (1999) Optimal temperature selection for mutation detection by denaturing HPLC and comparison to single-stranded conformation polymorphism and heteroduplex analysis. Clin. Chem. 45, 1133–1140.
Choy, Y. S., Dabora, S. L., Hall, F., et al. (1999) Superiority of denaturing high performance liquid chromatography over single-stranded conformation and conformation-sensitive gel electrophoresis for mutation detection in TSC2. Ann. Hum. Genet. 63, 383–391.
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Elanko, N., Jeffery, S. (2006). Mutation Analysis of PTPN11 in Noonan Syndrome by WAVE. In: Kearns-Jonker, M. (eds) Congenital Heart Disease. Methods in Molecular Medicine, vol 126. Humana Press. https://doi.org/10.1385/1-59745-088-X:97
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DOI: https://doi.org/10.1385/1-59745-088-X:97
Publisher Name: Humana Press
Print ISBN: 978-1-58829-375-6
Online ISBN: 978-1-59745-088-1
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