Abstract
Base excision sequence scanning (BESS) is used to detect and localise point mutations in mammalian genes. The cost of BESS can be significant in large-scale projects. however, due to the requirement for end-labelling of one of the two PCR primers. This is especially true when a fluorescent label is used for detection. If a universal label could be used for all of the PCR reactions, it would reduce the cost of this technique significantly. Here we describe a TP-BESS procedure where one fluorescence-labelled primer is used as a universal label for all the PCR reactions in BESS analysis. The universal label makes BESS financially more feasible for large-scale detection of single nucleotide polymorphisms (SNPs).
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Abbreviations
- BESS:
-
base excision sequence scanning
- IRD:
-
infrared fluorescence dye
- SNPs:
-
single nucleotide polymorphisms
- TP-BESS:
-
tailed primer base excision sequence scanning
References
Brieger J, Weidt EJ, Gansen K and Decker HJ (1999) Detection of novel germline mutation in the von Hippel-Lindau tumor-suppressor gene by fluorescence-labelled base excision sequence scanning (F-BESS). Clin Genet 56: 210–215.
Hawkins GA and Hoffman LM (1997) Base excision sequence scanning: a new method for rapid sequence scanning and mutation detection. Nature Biotechnology 15: 803–804.
Hawkins GA and Hoffman LM (1999) Rapid DNA mutation identification and fingerprinting using base excision sequence scanning. Electrophoresis 20: 1171–1176.
Meyers R, Maniatis T and Lerman L (1986) Detection and localization of single base changes by denaturing gradient gel electrophoresis. Methods Enzymol 155: 501–527.
Orita M, Iwahana H, Hayashi K and Sekiya T (1989) Detection of polymorphisms of human DNA by gel electrophoresis as single-strand conformation polymorphisms. Proc Natl Acad Sci USA 86: 2766–2770.
Wang D, Fan J, Siao C, Berno A, Young P, Sapolsky R, Ghandour G, Perkins N, Winchester E, Spencer J, Kruglyak L, Stein L, Hsie L, Topaloglou T, Hubbell E, Robinson E, Mittmann M, Morris M, Shen N, Kilburn D, Rioux J, Nusbaum C, Rozen S, Hudson T, Lipshutz R, Chee M and Lander E (1998) Large-scale identification, mapping, and genotyping of single-nucleotide polymorphisms in the human genome. Science 280: 1077–1082.
Yu K and Pauls KP (1994) Optimization of DNA-extraction and PCR procedures for random amplified polymorphic DNA (RAPD) analysis in plants. In: Griffin HG and Griffin AM (eds), PCR technology: current innovations, pp 193–200, CRC Press, Boca Raton, Florida.
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Yu, K., Haffner, M. & Poysa, V. Tailed primer base excision sequence scanning (TP-BESS) for detection of single nucleotide polymorphisms (SNPs). Plant Mol Biol Rep 19, 49–54 (2001). https://doi.org/10.1007/BF02824077
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DOI: https://doi.org/10.1007/BF02824077