Method for construction of normalized cDNA libraries

Details Method for construction of normalized cDNA libraries Method for construction of normalized cDNA libraries

1993-09-24

1996-01-09

Parr, Margaret Moskowitz

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435 6, 536 254, 935 80, C12P 1934

Patent

active

054828457

ABSTRACT:
This invention provides a method to normalize a directional cDNA library constructed in a vector that allows propagation in single-stranded circle form comprising: (a) propagating the directional cDNA library in single-stranded circles; (b) generating fragments complementary to the 3' noncoding sequence of the single-stranded circles in the library to produce partial duplexes; (c) purifying the partial duplexes; (d) melting and reassociating the purified partial duplexes to moderate Cot; and (e) purifying the unassociated single-stranded circles, thereby generating a normalized cDNA library.

REFERENCES:
Ko, M. S. H., (1990) An `equalized cDNA library` by the reassociation of short double-stranded cDNAs. Nucleic Acids Research, vol. 18, No. 19, pp. 5705-5711. (Exhibit B).
Patanjali, S. R., et al. (Mar. 1991) Construction of a uniform-abundance (normalized) cDNA library. Proc. Natl. Acad. Sci. U.S.A., 88:1943-1947. (Exhibit C).
NTIS Progress Report, Soares & Efstratiadis, Oct. 1992.

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