Competitive Oligonucleotide Single-Base Extension Combined with Mass Spectrometric Detection for Mutation Screening
| Autor: | G. Scott Higgins, Daniel P. Little, Hubert Köster |
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| Jazyk: | angličtina |
| Rok vydání: | 1997 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 23, Iss 4, Pp 710-714 (1997) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/97234st05 |
| Popis: | A rapid, robust and widely applicable mutation detection scheme not requiring radioactivity or gel-based detection is introduced. It is a single-tube, competitive oligonucleotide single-base extension (COSBE) reaction using a pair of primers with the 3′-terminal base complementary to either the normal or mutant allele. Upon hybridization and addition of a polymerase and the nucleotide triphosphate corresponding to the next base after the primer, only those primers properly annealed (i.e., no 3′-terminal mismatch) are extended; products are resolved by molecular weight shifts as determined by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (single-scan spectrum acquisition < |
| Databáze: | Directory of Open Access Journals |
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