Retrieval of DNA Sequences Present at an Extremely Low Frequency | Zendy

Alexandros Zafiropoulos | Zendy; G. Hatzidakis | Zendy; Lampros A. Mavrogiannis | Zendy; Apostolos Klinakis | Zendy; M. Kandilogiannaki | Zendy; Elias Krambovitis | Zendy

AI Assistant Blog Pricing

Home ZAIA Blog

Open Access

Retrieval of DNA Sequences Present at an Extremely Low Frequency

Author(s) -

Alexandros Zafiropoulos,

G. Hatzidakis,

Lampros A. Mavrogiannis,

Apostolos Klinakis,

M. Kandilogiannaki,

Elias Krambovitis

Publication year - 1997

Publication title -

biotechniques

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.617

H-Index - 131

eISSN - 1940-9818

pISSN - 0736-6205

DOI - 10.2144/97236rr02

Subject(s) - dna , biology , dna sequencing , genetics , population , microbiology and biotechnology , cloning (programming) , hybridization probe , point mutation , molecular cloning , nucleic acid thermodynamics , mutation , base sequence , gene , complementary dna , demography , sociology , computer science , programming language

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 x 10(3) and 1 in 28 x 10(3) 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 404,000-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.

The content you want is available to Zendy users.

Already have an account? Click here to sign in.

Having issues? You can contact us here

Accelerating Research