Abstract
Small insertions or deletions of nucleotides are common polymorphic variations in the human genome and can result in a predisposition to disease. However, high throughput methods for detecting these variations are limited. This report describes a method to detect this variation based on sequencing the boundaries of nucleotide alterations using the Pyrosequencing technique. This method can optimally detect up to 100 base pair nucleotide insertions and deletions, and also complicated genomic rearrangements.
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References
Ahmadian A, Gharizadeh B, Gustafsson AC, Sterky F, Nyren P, Uhlen M, Lundeberg J (2000) Single-nucleotide polymorphism analysis by pyrosequencing. Anal. Biochem. 280: 103–110.
Alderborn A, Kristofferson A, Hammerling U (2000) Determination of single-nucleotide polymorphisms by real-time pyrophosphate DNA sequencing. Genome Res. 10: 1249–1258.
Gupta PA, Putnam EA, Carmical SG, Kaitila I, Steinmann B, Child A, Danesino C, Metcalfe K, Berry SA, Chen E, Delorme CV, Thong MK, Ades LC, Milewicz DM (2002) Ten novel FBN2 mutations in congenital contractural arachnodactyly: delineation of the molecular pathogenesis and clinical phenotype. Hum. Mutat. 19: 39–48.
Kalinina O, Lebedeva I, Brown J, Silver J (1997) Nanoliter scale PCR with TaqMan detection. Nucl. Acids Res. 25: 1999–2004.
Little DP, Braun A, Darnhofer-Demar B, Frilling A, Li Y, McIver Jr. RT, Koster H (1997) Detection of RET proto-oncogene codon 634 mutations using mass spectrometry. J Mol. Med. 75: 745–750.
Nilsson M, Krejci K, Koch J, Kwiatkowski M, Gustavsson P, Landegren U (1997) Padlock probes reveal single-nucleotide differences, parent of origin and in situ distribution of centromeric sequences in human chromosomes 13 and 21. Nat. Genet. 16: 252–255.
Pourmand N, Elahi E, Davis RW, Ronaghi M (4-1-2002) Multiplex Pyrosequencing. Nucl. Acids Res. 30: e31.
Risch N, de Leon D, Ozelius L, Kramer P, Almasy L, Singer B, Fahn S, Breakefield X, Bressman S (1995) Genetic analysis of idiopathic torsion dystonia in Ashkenazi Jews and their recent descent from a small founder population. Nat. Genet. 9: 152–159.
Ronaghi M (2001) Pyrosequencing sheds light on DNA sequencing. Genome Res. 11: 3–11.
Syvanen AC (1999) From gels to chips: ‘minisequencing’ primer extension for analysis of point mutations and single nucleotide polymorphisms. Hum. Mutat. 13: 1–10.
Wight TN (2002) Versican: a versatile extracellular matrix proteoglycan in cell biology. Curr. Opin. Cell Biol. 14: 617–623.
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Guo, Dc., Qi, Y., He, R. et al. High throughput detection of small genomic insertions or deletions by Pyrosequencing. Biotechnology Letters 25, 1703–1707 (2003). https://doi.org/10.1023/A:1026090218031
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DOI: https://doi.org/10.1023/A:1026090218031