C - Chemistry – Metallurgy – 12 – N
Patent
C - Chemistry, Metallurgy
12
N
C12N 15/10 (2006.01) C07H 21/04 (2006.01)
Patent
CA 2526648
Disclosed herein is a method for synthesizing a desired nucleic acid sequence. The method comprises dividing the desired sequence into a plurality of partially overlapping segments; optimizing the melting temperatures of the overlapping regions of each segment to disfavor hybridization to the overlapping segments which are non-adjacent in the desired sequence; allowing the overlapping regions of single stranded segments which are adjacent to one another in the desired sequence to hybridize to one another under conditions which disfavor hybridization of non-adjacent segments; and filling in, ligating, or repairing the gaps between the overlapping regions, thereby forming a double-stranded DNA with the desired sequence. Also disclosed is a method for preventing errors in the synthesis of the nucleic acid sequence.
La présente invention a trait à un procédé permettant la synthèse d'une séquence d'acide nucléique souhaitée. Le procédé comprend la division de la séquence souhaitée en une pluralité de segments se chevauchant en partie ; l'optimisation des températures de fusion des régions de chevauchement de chaque segment pour défavoriser l'hybridation aux segments se chevauchant qui ne sont pas adjacents dans la séquence souhaitée ; permettant l'hybridation des unes aux autres des régions se chevauchant de segments à simple brin qui sont mutuellement adjacents dans des conditions qui défavorisent l'hybridation des segments non adjacents ; et le remplissage, la ligature, ou la réparation des espaces entre les régions se chevauchant, formant ainsi un ADN à double brin avec la séquence souhaitée. L'invention a également trait à un procédé permettant d'éviter des erreurs dans la synthèse de la séquence d'acide nucléique.
Hatfield G. Wesley
Lathrop Richard H.
Robic
The Regents Of The University Of California
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