Method for producing polymers
The invention relates to a method for producing polymers, in particular synthetic nucleic acid double strands of optional sequence, comprising the steps: (a) provision of a support having a surface area which contains a plurality of individual reaction areas, (b) location-resolved synthesis of nucleic acid fragments having in each case different base sequences in several of the individual reaction areas, and (c) detachment of the nucleic acid fragments from individual reaction areas.
1 . Method for synthesizing a minimal genome or a section thereof, characterized in that a plurality of oligomeric building blocks is synthesized on a support by parallel synthesis steps, is detached from the support and is brought into contact with one another to synthesize the minimal genome or section thereof.
2 . Method according to claim 1 characterized in that nucleic acid polymers selected from the group consisting of genes, minimal genes, gene clusters, chromosomes or sections thereof are synthesized and are brought into contact with one another to synthesize the minimal genome or section thereof.
3 . Method according to claim 1 characterized in that the minimal genome is a viral or bacterial genome.
4 . Method according to claim 1 characterized in that a double-stranded nucleic acid polymer with a length at least 100,000 bp is synthesized.
5 . Method according to claim 1 characterized in that the oligomeric building blocks are from 5 to 150, preferably 5 to 30 monomer units in length.
6 . Method according to claim 1 characterized in that in successive steps in each case partially complementary oligonucleotide building blocks are detached from the support and are brought into contact with one another or with the polymer intermediate under hybridization conditions.
7 . Method according to claim 1 for producing synthetic nucleic acid double strands, comprising the steps:
(a) provision of a support having a surface area which contains a plurality of individual reaction areas,
(b) location-resolved synthesis of nucleic acid fragments having in each case different base sequences in several of the individual reaction areas, and
(c) detachment of the nucleic acid fragments from individual reaction areas.
8 . Method according to claim 7 characterized in that the base sequences of the nucleic acid fragments synthesized in individual reaction areas are chosen such that they can assemble to form a nucleic acid double strand hybrid.
9 . Method according to claim 7 characterized in that the nucleic acid fragments according to step (c) are detached in one or more steps under conditions such that a plurality of the detached nucleic acid fragments assemble to form a nucleic acid double strand hybrid.
10 . Method according to claim 9 characterized in that several nucleic acid fragments which form one strand of the nucleic acid double strand hybrid are linked covalently to one another.
11 . Method according to claim 10 characterized in that the covalent linking includes treatment with ligase or/and filing in gaps in the strands using DNA polymerase.
12 . Method according to claim 7 characterized in that the sequence comprises at one or more positions recognition sequences for specific interaction with molecules such as proteins, nucleic acids, peptides, pharmaceuticals, saccharides, lipids, hormones, or/and organic compounds.
13 . Method according to claim 7 characterized in that the sequence of the nucleic acid double strands is a naturally occurring sequence, a not naturally occurring sequence or a combination of these two.
14 . Method according to claim 7 characterized in that the sequence is taken from a database, a sequencing experiment or a device for the integrated synthesis and analysis of polymers.
15 . Method according to claim 1 characterized in that the oligomeric building blocks are synthesized by location- or/and time-resolved illumination by means of a programmable light source matrix.
16 . Method according to claim 1 characterized in that a location- or/and time-resolved synthesis of the oligomeric building blocks takes place in a microfluidic reaction support having one or more fluidic reaction compartments and one or more reaction areas within a fluidic reaction compartment.
17 . Method according to claim 1 characterized in that the synthesis building blocks contain nucleotides occurring in nature, modified nucleotides or mixtures thereof
18 . Method according to claim 1 characterized in that modified synthesis building blocks are used for labelling and subsequent detection of the assembled nucleic acid double strands.
19 . Method according to claim 18 characterized in that the labelling groups used are molecules which are to be detected in a light-dependent manner.
20 . Use of a nucleic acid double strand produced according to the method according to claim 1 for therapeutic or pharmacological purposes.
21 . Use of a nucleic acid double strand produced according to the method according to claim 1 for diagnostic purposes.
22 . Use according to claim 20 comprising direct application to the intended purpose.
23 . Use according to claim 21 comprising direct application to the intended purpose.
24 . Use according to claim 20 comprising a conversion in effector cells.
25 . Use according to claim 21 comprising a conversion in effector cells.
26 . Use of a nucleic acid double strand produced according to the method according to claim 1 , where said nucleic acid double strand is stabilized, condensed or/and topologically manipulated during or following the combination and assembly in stages.
27 . Use according to claim 26 , where stabilization, condensation or/and topological manipulation is carried out by functional molecules such as histones or topoisomerases.
28 . Use of a nucleic acid double strand produced according to the method according to claim 1 as propagatable cloning vector, where the propagatable cloning vector may serve for transcription, expression of the transcribed sequence, and, where appropriate, production of expressed gene products in suitable target cells.