Method for the synthesis of dna sequences

C - Chemistry – Metallurgy – 12 – Q

Patent

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Details

C12Q 1/68 (2006.01) C07F 9/24 (2006.01) C07H 19/00 (2006.01) C07H 19/06 (2006.01) C07H 19/10 (2006.01) C07H 19/16 (2006.01) C07H 19/20 (2006.01) C07H 21/00 (2006.01) C07H 21/04 (2006.01) C12N 15/10 (2006.01) C12N 15/11 (2006.01) C07B 61/00 (2006.01)

Patent

CA 2447240

A method is disclosed for the direct synthesis of double stranded DNA molecules of a variety of sizes and with any desired sequence. The DNA molecule to be synthesis is logically broken up into smaller overlapping DNA segments. A maskless microarray synthesizer is used to make a DNA microarray on a substrate in which each element or feature of the array is populated by DNA of a one of the overlapping DNA segments. The DNA segments are released from the substrate and held under conditions favoring hybridization of DNA, under which conditions the segments will spontaneously hybridize together to form the desired DNA construct. This method makes possible the remote assembly of DNA sequence, through a process analogous to facsimile transmission of documents, since the information on DNA to be made can be transmitted remotely to an instrument which can then synthesize any needed DNA sequence from the information.

La présente invention concerne un procédé pour la synthèse directe de molécules d'ADN à double brin de différentes tailles et avec toute séquence souhaitée. On divise la molécule d'ADN dont on veut effectuer la synthèse en de plus petits segments d'ADN se chevauchant. On utilise un synthétiseur dépourvu de masque à jeu ordonné de microéchantillons pour constituer un jeu ordonné d'échantillons d'ADN sur un substrat dans lequel chaque élément ou composant du jeu est peuplé par l'ADN d'un parmi les segments d'ADN se chevauchant. On libère les segments d'ADN à partir du substrat et on les maintient dans des conditions favorisant l'hybridation d'ADN, dans lesquelles conditions les segments s'hybrideront ensemble de manière spontanée pour constituer la construction désirée. Ce procédé permet l'assemblage à distance d'une séquence d'ADN, grâce à un processus analogue à l'émission de télécopie de documents, étant donné que l'information concernant l'ADN à réaliser peut être transmise à distance vers un instrument qui peut effectuer la synthèse de toute séquence d'ADN requise à partir de l'information.

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