专利号:US-10501782-B1 优先权日:2014-09-05 标题:Cell-free synthesis of DNA by strand displacement 发明人:PORTER NEIL; ROTHWELL PAUL; EXTANCE JONATHAN 权利人:TOUCHLIGHT IP LTD 摘要:The present invention relates to an improved process for synthesis of DNA, RNA, proteins and like molecules, in particular cell-free enzymatic synthesis of DNA, preferably in large scale. The present invention relates to the synthesis of DNA using strand-displacement replication and the addition of nucleotides to the reaction mixture is controlled, thus controlling yield. The reaction mixture contains a starting amount of nucleotides, polymerase and DNA template, to which further nucleotides are supplied in a controlled manner.
专利号:US-6576447-B2 优先权日:2000-03-21 标 题:Method for synthesis of nucleic acids 发明人:TONOIKE HIROSHI 权利人:SHIMADZU CORP 摘要:An object of the present invention is to provide a novel method for suppressing the action of nucleic acid synthesis inhibitory substances and thereby amplifying a nucleic acid in a sample efficiently.According to the present invention, in a method for synthesis of nucleic acids to amplify an intended nucleic acid in a sample, the sample is brought in advance into contact with an insoluble polymer of a polyanion, a sulfated polymer or a sulfated polysaccharide, to remove nucleic acid synthesis inhibitory substances.
专利号:US-6767723-B2 优先权日:2000-04-27 标题 :Method for synthesis of nucleic acids 发明人:TONOIKE HIROSHI 权利人:SHIMADZU CORP 摘要:An object of the present invention is to provide a method of treatment and a method of storage that are useful in conducting a nucleic acid synthesis procedure capable of directly amplifying an intended nucleic acid in a living body-derived sample without purification steps.The present invention provides a method for synthesis of nucleic acids in which a living body-derived sample itself is mixed with a reaction solution for gene amplification and allowed to react, which method comprises treating the sample with a surfactant before the reaction to destruct solid components such as cells or bacterial bodies containing nucleic acids and uniformly disperse them in the sample liquid.
专利号:US-5514569-A 优先权日:1992-12-23 标 题:Method for enzymatic synthesis of oligonucleotides using phosphate precipitation 发明人:HYMAN EDWARD D 摘要:Enzymatic synthesis of a portion of an oligonucleotide is performed by a cycle of synthetic steps: (a) combining an oligonucleotide primer and a blocked nucleotide in a reaction mixture in the presence of a chain extending enzyme, such that a primer-blocked nucleotide product is formed, wherein the blocked nucleotide substrate comprises (i) a nucleotide to be added to form part of the defined sequence and (ii) a blocking group attached to the nucleotide effective to prevent the addition of more than one blocked nucleotide to the primer; and (b) removing the blocking group from the 3' end of the primer-blocked nucleotide product to form a primer-nucleotide product. Phosphate is generated in at least one synthetic step. A precipitate is formed in the cycle comprising phosphate and at least one precipitation cation. The precipitation of phosphate reduces its unfavorable effect on the method. Preferably, cycles of the method are repeated without intermediate purification of primer-nucleotide product or precursor. The precipitation cation may be a polyvalent elemental cation, spermine, or a cation which forms a poorly soluble salt with phosphate. Preferably, the chain extending enzyme is RNA Ligase, the blocked nucleotide is AppNp, and the blocking group is removed using a phosphatase. There is also provided a method for reducing the inhibitory effect of nucleoside 5'-monophosphate and of 3',5'-nucleoside diphosphate on phosphodiesterase I. This is accomplished by enzymatically converting these inhibitors to less inhibitory products.
专利号:US-6962780-B2 优先权日:2000-06-12 标 题 :Method for synthesis of nucleic acids 发明人:NAKAYAMA TOMOKO; NISHIMURA NAOYUKI 权利人:SHIMADZU CORP 摘要:The present invention is a method for synthesis of nucleic acids to amplify an intended nucleic acid in a region in which a GC content is rich, wherein a polyhydric alcohol and/or ammonium sulfate is present in an amplification reaction solution. According to the present invention, it is possible to amplify nucleic acids in a GC rich region efficiently and directly from a sample such as blood containing lots of PCR inhibitory substances without undergoing a process of isolating and purifying the nucleic acid, even though conducting PCR in the GC rich region tends to be difficult using conventional processes even if purified DNA is used.
专利号:US-2019323050-A1 优先权日:2016-12-21 标题 :Modulation of Enzymatic Polynucleotide Synthesis Using Chelated Divalent Cations 发明人:GRISWOLD JR KETTNER JOHN FREDERICK; LEE HOWON; CHURCH GEORGE M 权利人:HARVARD COLLEGE 摘要:Methods and apparatus of modulating polynucleotide synthesis are provided. The methods include delivering reagents comprising enzymes, nucleotides and ions to oligonucleotide primers wherein the reagents catalyze incorporation of the nucleotides to 3′ ends of the oligonucleotide primers, and modulating incorporation of the nucleotides to the 3′ ends of the oligonucleotide primers. Polynucleotide synthesis is modulated by modulating presence or absence of catalytic cation cofactors to provide sequence defined synthesis of polynucleotides. In certain embodiments, the polynucleotides encode information.
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