Process for conducting site-directed mutagenesis

Details Process for conducting site-directed mutagenesis Process for conducting site-directed mutagenesis

C - Chemistry, Metallurgy

12

N

C12N 15/10 (2006.01) C07H 21/04 (2006.01) C12N 9/38 (2006.01)

Patent

CA 2071892

2071892 9106643 PCTABS00005 The present invention relates to an approach to site-directed mutagenesis of double-stranded DNA. This approach involves the generation of structures capable of directing full-length complementary-strand synthesis of DNA. The structures are formed following heat denaturation and cooling of linear DNA templates in the presence of what is a ''closing'' oligonucleotide. This closing oligonucleotide is a nucleotide sequence having at least part of its sequence complementary to at least one of the free ends of the DNA template. The closing oligonucleotide functions as an agent for circularization of a DNA strand and generation of a primer-circular template structure suitable for polymerase-dependent full-length complementary-strand synthesis and ligation into a covalently-closed heteroduplex DNA molecule.

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