专利号:US-2023235372-A1 优先权日:2020-06-12 标题:Ab-initio, template-independent synthesis of nucleic acids using thermostable enzymes 发明人:RANDRIANJATOVO-GBALOU IRINA; SAID AHMED; RAHIER RENAUD 权利人:SYNHELIX 摘要:The invention relates to the field of nucleic acid synthesis or sequencing, more specifically to methods for ab-initio synthesis of nucleic acids, comprising contacting a nucleotide with a free 3′-hydroxyl group, with at least one nucleoside triphosphate, or a combination of nucleoside triphosphates, in the presence of an archaeal DNA primase or a functionally active fragment and/or variant thereof, thereby covalently binding said nucleoside triphosphate to the free 3′-hydroxyl group of the nucleotide. It also relates to isolated functionally active fragments of archaeal DNA primases which are capable of both ab-initio single-stranded nucleic acid synthesis activity and template-independent terminal nucleotidyl transferase activity.
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-7344863-B2 优先权日:1998-03-13 标题:Methods for producing polypeptides through enhanced synthesis of encoding nucleic acid molecules 发明人:LI WU-BO; JESSEE JOEL A; SCHUSTER DAVID; XIA JIULIN; GEBEYEHU GULILAT 权利人:INVITROGEN CORP 摘要:The present invention is directed to compositions and methods for enhancing synthesis of nucleic acid molecules, particularly GC-rich nucleic acid molecules. Specifically, the invention provides compositions comprising one or more nitrogen-containing organic compounds having a formula selected from the group consisting of formula I and formula II (or salts or derivatives thereof), preferably 4-methylmorpholine N-oxide or betaine (carboxymethyltrimethylammonium), and further comprising one or more compounds selected from the group consisting of proline and an N-alkylimidazole compound, and more preferably proline, 1-methylimidazole or 4-methylimidazole. The invention further relates to methods for enhanced, high-fidelity synthesis of nucleic acid molecules, including via amplification (particularly PCR), reverse transcription, and sequencing methods. The invention also relates to nucleic acid molecules synthesized by these methods, to fragments or derivatives thereof, and to vectors and host cells comprising such nucleic acid molecules, fragments, or derivatives. The invention also relates to kits for synthesizing, amplifying, reverse transcribing or sequencing nucleic acid molecules comprising one or more of the compositions of the invention.
专利号:US-12421628-B2 优先权日:2018-07-23 标题:Massively parallel enzymatic synthesis of nucleic acid strands 发明人:HORGAN ADRIAN; GODRON XAVIER; YBERT THOMAS; NICOL ROBERT 权利人:DNA SCRIPT 摘要:The invention is directed to methods for massively parallel template-free enzymatic synthesis of a plurality of different polynucleotides of predetermined sequences. In one aspect, methods of the invention employ large scale arrays of reaction sites each associated with at least one working electrode for controlling deprotection and deblocking steps at predetermined user selected sites. In another aspect, the invention provides template-free enzymatic synthesis with proofreading, wherein completed polynucleotides at predetermined reaction sites are sequenced using a sequencing by synthesis technique, particularly employing electrochemically labile blocking groups.
专利号:US-5945312-A 优先权日:1996-04-15 标 题 :Synthesis of fluorophore-labeled DNA 发明人:GOODMAN MYRON F; REHA-KRANTZ LINDA J 权利人:UNIV SOUTHERN CALIFORNIA; UNIV ALBERTA 摘要:Sequencing methods and methods for synthesizing DNA probes using mutant bacteriophage T4 DNA polymerases which have increased ability to incorporate modified nucleotides for the synthesis of long or short chains of complementary, modified, e.g., fluorophore-labeled DNA. In general, the mutant T4 DNA polymerases retain 3'→5' exonuclease activity; hence, reduction or elimination of 3'→5' exonuclease activity is not a prerequisite for efficient synthesis of a complementary fluorophore-labeled or other modified DNA. In fact, retention of 3'→5' exonuclease activity increases accuracy of DNA replication, because these exonucleases proofread or edit the product of DNA replication.
专利号:US-8058414-B2 优先权日:2008-04-29 标题 :Unnatural polymerase substrates that can sustain enzymatic synthesis of double stranded nucleic acids from a nucleic acid template and methods of use 发明人:MENCHEN STEVEN; ROSENBLUM BARNETT; KENNEY PAUL; KHAN SHOEB; MA ZHAOCHUN; CHEN JER-KANG; LAM JOE Y; HUANG BOLI 权利人:MENCHEN STEVEN; ROSENBLUM BARNETT; KENNEY PAUL; KHAN SHOEB; MA ZHAOCHUN; CHEN JER-KANG; LAM JOE Y; HUANG BOLI; LIFE TECHNOLOGIES CORP 摘要:Nucleotide analogs that can sustain the enzymatic synthesis of double-stranded nucleic acid from a nucleic template are described. The nucleotide analogs include: (i) a base selected from the group consisting of adenine, guanine, cytosine, thymine, uracil and their analogs; (ii) a label attached to the base or analog of the base via a cleavable linker; (iii) a deoxyribose; and (iv) one or more phosphate groups. The linker and/or the label inhibits template directed polymerase incorporation of a further nucleotide substrate onto an extended primer strand. In addition, cleavage of the linker leaves a residue attached to the base which is not present in the natural nucleotide and which does not inhibit extension of the primer strand. The nucleotide analogs can therefore be used as reversible terminators in sequencing by synthesis methods without blocking the 3′ hydroxyl group. Methods of sequencing DNA using the substrates are also described.
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