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Oligonucleotide-directed mutagenesis: a sequence-based screening.

A Duilio·F Cimino·T Russo
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Abstract

A procedure to screen for mutant clones obtained by oligonucleotide-directed mutagenesis has been developed. It is based on the preparation of phage containing supernatants from a number (100 or more) of randomly chosen mutagenized M13 plaques. Aliquots from these supernatants are mixed to obtain pools, each containing 10 phages. Heterogeneous single-stranded DNA is prepared from these pools and used as template in a "single letter" sequence according to the dideoxy chain terminator method. Thus, the pool(s) containing the mutated sequence and the mutated sequence itself is identified by sequencing the single-stranded DNAs of the 10 phages present in the selected pool.

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AnimalsBase SequenceClone CellsCodonMutationOligonucleotidesRNA, Transfer, ProJournal ArticleResearch Support, Non-U.S. Gov't

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A Duilio, F Cimino, T Russo. 1988-11-01. Oligonucleotide-directed mutagenesis: a sequence-based screening.. https://doi.org/10.1016/0003-2697(88)90064-4

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