Abstract
Small alterations in DNA sequence of genomic DNA lead to many human diseases, such as cancer, diabetes, heart disease, atherosclerosis, cystic fibrosis, Alzheimer disease, Duchenne muscular dystrophy, and various thalassemias. These alterations in DNA sequence include many types of mutations and polymorphisms, such as substitutions of one or several nucleotides, deletions or insertions of some larger sequences, differences in variable number of tandem repeats (VNTR), and the genomic instability of microsatellite repeat (1). The diagnosis of human diseases by DNA polymorphism analysis has very important applications in the fields of genetic and medical research, clinical chemistry and forensic science. Because a large portion of sequence variations in the human genome is caused by single base changes, any method used to detect mutations or polymorphisms must be capable of detecting single-base substitutions.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kaplan, J. C., and Delpech, M. (1993) Biologie Moleculaire et Medicine, Flammarion, Paris.
Righetti, P. G., ed. (1996) Capillary Electrophoresis in Analytical Biotechnology, CRC Press, Boca Raton, pp. 431–437.
Wang, Y., Hung, S. C., Linn, J.F., Steiner, G., Glazer, A. N., Sidransky, D., and Mathies, R. A. (1997) Microsatellite-based cancer detection using capillary electrophoresis and energy-transfer fluorescent primers. Electrophoresis 18, 1742–1749.
Effenhauser, C. S., Bruin, G. J. M., and Paulus, A. (1997) Integrated chip-based capillary electrophoresis. Electrophoresis 18, 2203–2213.
Righetti, P. G., Gelfi, C., Perego, M., Stoyanov, A. V., and Bossi, A. (1997) Capillary zone electrophoresis of oligonucleotides and peptides in isoelectric buffers: theory and methodology. Electrophoresis 18, 2145–2153.
Righetti, P. G. and Gelfi, C. (1996) Capillary electrophoresis of DNA, in Capillary Electrophoresis in Analytical Biotechnology (Righetti, P. G., ed.), CRC Press, Boca Raton, pp. 431–476.
Heiger, D. N., Cohen, A. S., and Karger, B. L. (1990) Separation of DNA restriction fragments by high performance capillary electrophoresis with low and zero cross-linked polyacrylamide using continuous and pulsed electric fields. J. Chromatogr. 516, 33–44.
Gelfi, C., Orsi, A., Leoncini, F., and Righetti, P. G. (1995) Fluidified polyacrylamides as molecular sieves in capillary zone electrophoresis of DNA fragments. J. Chromatogr. A 689, 97–107.
Simò-Alfonso, E., Gelfi, C., Sebastiano, R., Citterio, A., and Righetti, P. G. (1996) Novel acrylamido monomers with higher hydrophilicity and improved hydrolytic stability. I: synthetic route and product characterization. Electrophoresis 17, 723–731.
Simò-Alfonso, E., Gelfi, C., Sebastiano, R., Citterio, A., and Righetti, P. G. (1996) Novel acrylamido monomers with higher hydrophilicity and improved hydrolytic stability. II: properties of N-acryloyl amino propanol. Electrophoresis 17, 732–737.
Gelfi, C., Simò-Alfonso, E., Sebastiano, R., Citterio, A., and Righetti, P. G. (1996) Novel acrylamido monomers with higher hydrophilicity and improved hydrolytic stability. III: DNA separations by capillary electrophoresis in poly(N-acryloyl amino propanol). Electrophoresis 17, 738–743.
Simò-Alfonso, E., Gelfi, C., Lucisano, M., and Righetti, P. G. (1996) Performance of a series of novel N-substituted acrylamides in capillary electrophoresis of DNA fragments. J. Chromatogr. A 756, 255–262.
Gelfi, C., Perego, M., Libbra, F., and Righetti, P. G. (1996) Comparison of the behaviour of N-substituted acrylamides and celluloses on double-stranded DNA separations by capillary electrophoresis at 25° and 60°C. Electrophoresis 17, 1342–1347.
Myers, R. M., Maniatis, T., and Lerman, L. (1987) Detection and localization of single base changes by denaturing gradient gel electrophoresis. Methods Enzymol. 155, 501–527.
Lerman, L. S. and Silverstein, K. (1987) Computational simulation of DNA melting and its application to denaturing gradient gel electrophoresis. Methods Enzymol. 155, 482–501.
Bello, M. S., Levin, E. I., and Righetti, P. G. (1993) Computer-assisted determination of the inner temperature and peak correction for capillary electrophoresis. J. Chromatogr. 652, 329–336.
Kälin, A., Dörk, T., Bozon, D., and Tümmler, B. (1992) A novel frame-shift mutation in exon 4 of the cystic fibrosis gene (435insA) demonstrates the ambiguity of restriction analysis for mutation screening. Hum. Mol. Genet. 1, 545–546.
Guillermit, H., Fanen, P., and Ferenc, C. (1990) A 3′ splice site consensus sequence mutation in the cystic fibrosis gene. Hum. Genet. 85, 450–453.
Kerem, B., Zielenski, J., Markiewicz, D., Bozon, D., Gazit, E., Jahaf, J., et al. (1990) Identification of mutations in regions corresponding to the two putative nucleotide (ATP)-binding folds of the cystic fibrosis gene. Proc. Natl. Acad. Sci. USA 87, 8447–8451.
Devoto, M., Ronchetto, P., Fanen, P., Telleria Orriolis, J. J., Romeo, G., Goossens, M., et al. (1991) Screening for non-deltaF508 mutations in five exons of the cystic fibrosis transmembrane conductance regulator gene in Italy. Am. J. Hum. Genet. 48, 1127–1132.
Zielenski, J., Rozmahel, R., Bozon, D., Kerem, B., Grzelczak, Z., Riordan, J., et al. (1991) Genomic DNA sequence of the cystic fibrosis transmembrane conductance regulator gene. Genomics 10, 214–228.
Brancolini, V., Cremonesi, L., Belloni, E., Pappalardo, E., Bordoni, R., Seia, M., et al. (1995) Search for mutations in pancreatic sufficient cystic fibrosis Italian patients: detection of 90% of molecular defects and identification of three novel mutations. Hum. Genet. 96, 312–318.
Gelfi, C., Righetti, P. G., Cremonesi, L., and Ferrari, M. (1994) Detection of point mutations by capillary electrophoresis in liquid polymers in temporal thermal gradients. Electrophoresis 15, 1506–1511.
Gelfi, C., Cremonesi, L., Ferrari, M., and Righetti, P. G. (1998) Temperature-programmed capillary electrophoresis for detection of DNA point mutations. BioTechniques 21, 926–932.
Gelfi, C., Righetti, P. G., Travi, M., and Fattore, S. (1997) Temperature programmed capillary electrophoresis for the analysis of high-melting point mutants in thalassemias. Electrophoresis 18, 724–731.
Cremonesi, L., Firpo, S., Ferrari, M., Righetti, P. G., and Gelfi, C. (1997) Double-gradient DGGE for optimized detection of DNA point mutations. BioTechniques 22, 326–330.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2001 Humana Press Inc.
About this protocol
Cite this protocol
Gelfi, C., Cremoresi, L., Ferrari, M., Righetti, P.G. (2001). Point Mutation Detection by Temperature-Programmed Capillary Electrophoresis. In: Mitchelson, K.R., Cheng, J. (eds) Capillary Electrophoresis of Nucleic Acids. Methods in Molecular Biology™, vol 163. Humana Press. https://doi.org/10.1385/1-59259-116-7:73
Download citation
DOI: https://doi.org/10.1385/1-59259-116-7:73
Publisher Name: Humana Press
Print ISBN: 978-0-89603-765-6
Online ISBN: 978-1-59259-116-9
eBook Packages: Springer Protocols