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 directly converting digital target sequences into nucleic acids comprising the steps
(a) providing a digital target sequence,
(b) fragmenting the digital target sequence into suitable oligomeric building blocks,
(c) synthesizing said oligomeric building blocks on at least one support by parallel synthesis steps,
(d) detaching the oligomeric building blocks from the support,
(e) bringing the oligomeric building blocks into contact with one another, and
(f) linking the phosphodiester backbone enzymatically.
2 . Method according to claim 1 , wherein the digital target sequence/nucleic acid is selected from the group consisting of genes or sections thereof, gene clusters or sections thereof, chromosomes or sections thereof, or viral or bacterial genomes or sections thereof
3 . Method according to claim 1 , wherein the digital target sequence of step (a) describes functional genetic elements out of the group consisting of genes or sections thereof, gene clusters or sections thereof, chromosomes or sections thereof, or viral or bacterial genomes or sections thereof; or variations thereof
4 . Method according to claim 1 , wherein the conversion of the digital target sequence into oligomeric building blocks includes variation of the nucleic acids.
5 . Method according to claim 1 , wherein the digital target sequence/nucleic acid is a nucleic acid double strand.
6 . Method according to claim 5 , wherein the digital target sequence/nucleic acid is a double-stranded nucleic acid of at least 300 bp in length.
7 . Method according to claim 1 , wherein the digital target sequence derives from a database.
8 . Method according to claim 1 , wherein suitable oligomeric building blocks are generated taking into account biochemical and functional parameters in the fragmenting step (b).
9 . Method according to claim 1 , wherein the building blocks are chosen such that they can assemble to form a nucleic acid double strand hybrid.
10 . Method according to claim 1 , wherein an algorithm makes out suitable overlapping regions in the fragmenting step (b).
11 . Method according to claim 1 , wherein the oligomeric building blocks are from 5-150 monomer units in length.
12 . Method according to claim 1 , wherein each oligomeric building block is synthesized on a different area of a common support (step (c)).
13 . Method according to claim 1 , wherein the synthesis of the building blocks (step (c)) is carried out in a location or/and time-resolved synthesis process.
14 . Method according to claim 1 , wherein the synthesis step (c), the detachment step (d) and the contacting step (e) is carried out within one compartment.
15 . Method according to claim 1 , wherein step (c) is carried out in a microfluidic reaction support having one or more fluidic reaction compartments and one or more reaction areas within a fluidic reaction compartment.
16 . Method according to claim 1 , wherein in step (d) in each ease partially complementary building blocks are detached from the support and are brought into contact with one another or with the polymer intermediate under hybridization conditions.
17 . Method according to claim 1 , wherein the oligomeric building blocks are detached in one or more steps under conditions such that a plurality of detached nucleic acid fragments assemble to form a nucleic acid double strand hybrid.
18 . Method according to claim 1 , wherein step (f) includes treatment with ligase.
19 . Method according to claim 1 , wherein possible gaps in the strands after hybridization are filled using polymerase.
20 . Method for generating functionally integrated DNA molecules, comprising the steps
(a) providing digital nucleic acid sequences of several functional elements,
(b) integrating the functional elements into a digital DNA molecule,
(c) fragmenting the digital DNA molecule into suitable oligomeric building blocks,
(d) synthesizing said oligomeric building blocks on at least one support by parallel synthesis steps,
(e) detaching the oligomeric building blocks from the support,
(f) bringing the oligomeric building blocks into contact with one another, and
(g) linking the phosphodiester backbone enzymatically to form the integrated DNA molecule.
21 . Method of claim 20 , wherein the functional element is selected from the group consisting of genes, part of genes, regulatory elements, viral packaging signals or viral vectors.
22 . Method according to claim 20 , wherein the digital DNA molecule derives from a database.
23 . Method according to claim 20 , wherein suitable oligomeric building blocks are generated taking into account biochemical and functional parameters in the fragmenting step (c).
24 . Method according to claim 20 , wherein the building blocks are chosen such that they can assemble to form a nucleic acid double strand hybrid.
25 . Method according to claim 20 , wherein an algorithm makes out suitable overlapping regions in the fragmenting step (b).
26 . Method according to claim 20 , wherein the oligomeric building blocks are from 5-150 monomer units in length.
27 . Method according to claim 20 , wherein each oligomeric building block is synthesized on a different area of a common support (step (d)).
28 . Method according to claim 20 , wherein the synthesis of the building blocks (step (d)) is carried out in a location or/and time-resolved synthesis process.
29 . Method according to claim 20 , wherein step (c) is carried out in a microfluidic reaction support having one or more fluidic reaction compartments and one or more reaction areas within a fluidic reaction compartment.
30 . Method according to claim 20 , wherein in step (e) in each case partially complementary building blocks are detached from the support and are brought into contact with one another or with the polymer intermediate under hybridization conditions.
31 . Method according to claim 20 , wherein the oligomeric building blocks are detached in one or more steps under conditions such that a plurality of detached nucleic acid fragments assemble to form a nucleic acid double strand hybrid.
32 . Method according to claim 20 , wherein step (g) includes treatment with ligase.
33 . Method according to claim 20 , wherein possible gaps in the strands after hybridization are filled using polymerase.