Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1994-01-19
1996-05-07
Jones, W. Gary
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
435 6, 435 5, 536 2433, 935 78, C12P 1934, C12Q 168, C12Q 170, C07H 2104
Patent
active
055145680
ABSTRACT:
The invention is directed to a method of introducing at least one predetermined change in a nucleic acid sequence of a double-strand DNA. The method includes: (a) providing a first primer and a second primer capable of directing said predetermined change in said nucleic acid sequence, said first and second primers comprising a nucleic acid sequence substantially complementary to said double-stranded DNA so as to allow hybridization, a class IIS restriction enzyme recognition sequence and cleavage sites; (b) hybridizing said first and second primers to opposite strands of said double-stranded DNA to form a first pair of primer-templates oriented in opposite directions; (c) extending said first pair of primer-templates to create double-stranded molecules; (d) hybridizing said first and second primers at least once to said double-stranded molecules to form a second pair of primer-templates; (e) extending said second pair of primer-templates to produce double-stranded linear molecules terminating with class IIS restriction enzyme recognition sequences; and (f) restricting said double-stranded linear molecules with a class IIS restriction enzyme to form restricted linear molecules containing said change in said nucleic acid sequence. Also provided is a method of producing at least two changes located at one or more positions within a nucleic acid sequence. Synthetic primers for producing such changes are provided as well.
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Arthur Lisa
Eli Lilly and Company
Jones W. Gary
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