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Deletion mutagenesis in M13 by polymerase chain reaction using universal sequencing primers



Analytical biochemistry 188 (1990), S.255-258
ISSN: 0003-2697
Fraunhofer ITA ( ITEM) ()
DNA; DNA sequences; mutagenesis; PCR; polymerase chain reaction; serine rich protein

A simple procedure is described for the efficient deletion of large DNA sequences. The method involves a combination of oligonucleotide-directed mutagenesis in bacteriophage M13 and amplification of the mutagenized product by polymerase chain reaction. In contrast to other protocols employing polymerase chain reaction, synthesis of only one specific primer is required. The efficiency of heteroduplex formation between mutagenic primers directing large deletions and singlestranded template is discussed.