Retrieval of DNA Sequences Present at an Extremely Low Frequency
| Autor: | A. Zafiropoulos, G. Hatzidakis, L. Mavrogiannis, A. Klinakis, M. Kandilogiannaki, E. Krambovitis |
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| Jazyk: | angličtina |
| Rok vydání: | 1997 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 23, Iss 6, Pp 1104-1109 (1997) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/97236rr02 |
| Popis: | We describe a method for retrieving sequences with one or two point mutations of a given target sequence, which are present in a DNA population at a frequency of 1 in 466 × 103 and 1 in 28 × 103 molecules, respectively. By stringent hybridization to a stable, chemically immobilized probe, a large excess of unrelated fragments is removed, and the bound sequences are dissociated and amplified. By repeating the hybridization-amplification cycles twice, we achieved an estimated enrichment of 404000-fold and 1612-fold, respectively, which was confirmed by cloning the resultant products and sequencing 35 clones. This procedure can be applied to retrieve mutated sequences that exist at an extremely low frequency in a DNA population. |
| Databáze: | Directory of Open Access Journals |
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