Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1993-09-02
1997-10-28
Guzo, David
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
4351721, C12P 1934
Patent
active
056817266
ABSTRACT:
The present invention provides an improved method for the synthesis of double stranded DNA, particularly complementary DNA for the construction of directional complementary DNA libraries. The method comprises synthesizing a first strand of DNA complementary to a selected RNA or DNA template by contacting with the template a linker/primer comprising a selected restriction site, a suitable RNA or DNA dependent DNA polymerase, and substrates comprising a deoxyribonucleotide triphosphate analog. The linker/primer and deoxyribonucleotide triphosphate analog are selected such that incorporation of the nucleotide analog in the first strand substantially protects the double stranded DNA from cleavage, under conditions sufficient to cleave or substantially cleave the linker/primer, at the selected restriction site.
REFERENCES:
Schreiber et al. "Selective Protection of in vitro Synthesized cDNA Against Nucleases by Incorporation of Phosphorothioate-Analogues" Nucleic Acids Research, vol. 13, No. 21, pp. 7663-7672, 1985.
Han et al. "Isolation of Full-Length Putative Rat Lysophospholipase cDNA Using Improved Methods for mRNA Isolation and cDNA Cloning", Biochemistry, vol. 26, pp. 1617-1625, 1987.
Vosberg et al. "Effect of Deoxynucleoside Phosphorothioates Incorporated in DNA on Cleavage . . . " JBC, vol. 257, pp. 6595-6599 (1982).
Krawinkel et al. (1986) "Rapid Synthesis of cDNA of Cloning," NAR, vol. 14, p. 1913.
Maniatis et al. (1982) "Molecular Cloning" CSH (New York) pp. 197-198.
Gruenbaum et al. (1981) "Restriction Enzyme Digestion of Hemimethylated DNA" NAR, vol. 9, pp. 2509-2515.
Nelson et al (1987). "The Effect of Site-Specific Methylation on Restriction-Modification Enzymes." NAR, vol. 15, pp. r219-r230.
Hansen Connie Jo
Huse William David
Guzo David
Halluin Albert P.
Stratagene
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