Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Preparing compound containing saccharide radical
Patent
1989-08-23
1992-07-14
Moskowitz, Margaret
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Preparing compound containing saccharide radical
435 6, 4351723, 435810, 436 94, 436508, 536 27, 536 28, C12P 1934, C12Q 168, C07H 1512, C07H 1700
Patent
active
051302381
ABSTRACT:
This invention relates to an improved process for amplifying a specific nucleic acid sequence. The process involves synthesizing single-stranded RNA, single-stranded DNA and Double-stranded DNA. The single-stranded RNA is a first template for a first primer, the single-stranded DNA is a second template for a second primer, and the double stranded DNA is a third template for synthesis of a plurality of copies of the first template. A sequence of the first primer or the second primer is complementary to a sequence of the specific nucleic acid and a sequence of the first primer or the second primer is homologous to a sequence of the specific nucleic acid. The improvement of the amplification process involves the addition of DMSO alone or in combination with BSA, which improves the specificity and efficiency of the amplification. The amplification process may be used to increase the quantity of a specific nucleic acid sequence to allow detection, or to increase the purity of a specific nucleic acid sequence as a substitute for conventional cloning methodology.
REFERENCES:
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patent: 4683202 (1987-07-01), Mullis
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Scapes Protein Purification (1982) Springer-Verlag New York Inc. NY NY p. 197.
Bethesda Research Laboratories Catalogue of 1988 Bethesda, M.D. pp. 30 and 37.
Gelfand et al. "Principles and Applications for DNA Amplification", PCR Technologies, Stockton Press, New York (1989).
Scharf et al. "Direct Cloning and Sequence Analysis of Enzymatically Amplified Genomic Sequences", Science, 233:1076-1078 (1976).
Maniatis et al. Molecular Cloning--A Laboratory manual (Cold Spring Harbor Laboratory 1982).
Okayama et al. "High-Efficiency Cloning of Full-Length cDNA", Mol. Cel. Biol., 2:161 (1982).
Gubler et al. "A Simple and Very Efficient Method for Generating cDNA Libraries", Gene, 25:263-269 (1983).
Polites et al. "A Step-Wise Protocol for cDNA Synthesis", Bio Techniques 4:514 (1986).
Davey Cheryl
Henderson Graham
Malek Lawrence T.
Sooknanan Roy
Cangene Corporation
Chambers Scott A.
Moskowitz Margaret
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