Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1984-02-17
1989-06-27
Wiseman, Thomas G.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
4351723, 435 6, 536 27, 935 1, 935 22, 935 77, C12P 1932, C12N 1500, C07H 1512
Patent
active
048430030
ABSTRACT:
A process is provided for producing an ordered series of cloned, circular, DNA molecules containing shortened target DNA segments derived from a long target DNA segment, which are suitable for use in determining the nucleotide sequence of the long target DNA segment, or for targeting specific regions within the target DNA segment. The process includes producing, by molecular cloning, a plurality of double-stranded recombinant DNA molecules each containing: (i) vector DNA; (ii) a sequencing primer binding site; and, (iii) a DNA region having unique endonuclease sites and a long target DNA segment. The sequencing primer binding site is spaced from the long target DNA segment by at least a portion of said DNA region having the unique endonuclease sites. The plurality of double-stranded circular recombinant DNA molecules are cleared using two restriction endonucleases. The cleavage occurs in the portion of the DNA having unique endonuclease sites lying between the long target DNA segment and the sequencing primer binding site. The cleavage and, if necessary, subsequent processing steps, produces double-stranded linear DNA molecules having an end containing a long target DNa segment that is susceptible to exonuclease digestion and a sequencing primer binding site end that is not susceptible to exonuclease digestio
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Gelinas Richard E.
Henikoff Steven
Fred Hutchinson Cancer Research Center
Mays Thomas D.
Wiseman Thomas G.
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