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1. (WO2018111914) METHODS FOR ASSEMBLING DNA MOLECULES
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Pub. No.: WO/2018/111914 International Application No.: PCT/US2017/065877
Publication Date: 21.06.2018 International Filing Date: 12.12.2017
IPC:
C12N 15/09 (2006.01) ,C12N 15/10 (2006.01) ,C12P 19/34 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
10
Processes for the isolation, preparation or purification of DNA or RNA
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
P
FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
19
Preparation of compounds containing saccharide radicals
26
Preparation of nitrogen-containing carbohydrates
28
N-glycosides
30
Nucleotides
34
Polynucleotides, e.g. nucleic acids, oligoribonucleotides
Applicants:
SYNTHETIC GENOMICS, INC. [US/US]; 11149 North Torrey Pines Road, Suite 100 La Jolla, CA 92037, US
Inventors:
GILL, John, E.; US
GIBSON, Daniel, G.; US
FU, Lixia; US
Agent:
HAILE, Lisa, A.; US
HIERHOLZER, Matthew, J.; US
LIMBACH, Alan, A.; US
HWANG, Pamela, E.; US
JACKSON, Blake; US
Priority Data:
62/434,30014.12.2016US
Title (EN) METHODS FOR ASSEMBLING DNA MOLECULES
(FR) PROCÉDÉS D’ASSEMBLAGE DE MOLÉCULES D’ADN
Abstract:
(EN) The invention provides compositions and methods for assembling a DNA molecule having a desired sequence. The methods involve contacting a DNA polymerase, dNTPs, and a plurality of pairs of oligonucleotides. The oligonucleotides of a pair have a portion of the desired sequence, and an internal sequence that overlaps and is complementary to an internal sequence of the other oligonucleotide of the pair, and, when arranged in order, they have at least a portion of the desired sequence. The oligonucleotides also have a 3' or a 5' primer binding sequence having a binding site for a primer. The oligonucleotides that correspond to the end oligonucleotides of the desired sequence also have a universal 3' flanking sequence and a universal 5' flanking sequence, respectively. The methods involve performing a first amplification reaction on the plurality of pairs of oligonucleotides; removing the 3' and 5' primer binding sequences from the plurality of pairs of oligonucleotides; and subjecting the plurality of pairs of oligonucleotides to an assembly reaction to thereby assemble the dsDNA molecule having the desired sequence.
(FR) La présente invention concerne des compositions et des procédés d’assemblage d’une molécule d’ADN présentant une séquence souhaitée. Les procédés consistent à mettre en contact une ADN polymérase, des dNTP, et une pluralité de paires d’oligonucléotides. Les oligonucléotides d’une paire présentent une partie de la séquence souhaitée, et une séquence interne qui chevauche et qui est complémentaire d’une séquence interne de l’autre oligonucléotide de la paire, et, lorsque disposées dans l’ordre, elles possèdent au moins une partie de la séquence souhaitée. Les oligonucléotides possèdent également une séquence de liaison d’amorce (3’) ou (5’) ayant un site de liaison pour une amorce. Les oligonucléotides qui correspondent aux oligonucléotides terminaux de la séquence souhaitée possèdent respectivement également une séquence flanquante (3’) universelle et une séquence flanquante (5’) universelle. Les procédés consistent à effectuer une première réaction d’amplification sur la pluralité de paires d’oligonucléotides ; à éliminer les séquences de liaison d’amorce (3’) et (5’) de la pluralité des paires d’oligonucléotides ; et à soumettre la pluralité des paires d’oligonucléotides à une réaction d’assemblage pour assembler ainsi la molécule d’ADNdb possédant la séquence souhaitée.
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Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (EPO) (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: English (EN)
Filing Language: English (EN)