Base composition-independent hybridization in tetramethylammonium chloride: a method for oligonucleotide screening of highly complex gene libraries. | Zendy

William I. Wood | Zendy; Jane Gitschier | Zendy; L A Lasky | Zendy; Richard M. Lawn | Zendy

Open Access

Base composition-independent hybridization in tetramethylammonium chloride: a method for oligonucleotide screening of highly complex gene libraries.

Author(s) -

William I. Wood,

Jane Gitschier,

L A Lasky,

Richard M. Lawn

Publication year - 1985

Publication title -

proceedings of the national academy of sciences

Language(s) - English

Resource type - Journals

eISSN - 1091-6490

pISSN - 0027-8424

DOI - 10.1073/pnas.82.6.1585

Subject(s) - oligonucleotide , tetramethylammonium , base (topology) , mutagenesis , oligonucleotide synthesis , composition (language) , biology , function (biology) , base pair , oligomer restriction , computational biology , gene , chemistry , microbiology and biotechnology , combinatorial chemistry , biochemistry , genetics , mutation , organic chemistry , ion , mathematical analysis , linguistics , philosophy , mathematics

An oligonucleotide hybridization procedure has been developed that eliminates the preferential melting of A X T versus G X C base pairs, allowing the stringency of the hybridization to be controlled as a function of probe length only. This technique, which uses tetramethylammonium chloride, is especially helpful whenever a highly complex library is screened with a pool of oligonucleotide probes, which usually vary widely in base composition. The procedure can also be applied advantageously whenever an exact match to an oligonucleotide probe is desired, such as in screening for clones having as little as a single-base alteration generated by in vitro mutagenesis.

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