DE19819735A1 - Device and method for producing an arrangement of chain molecules on a carrier material - Google Patents

Abstract

The invention relates to a method, a device and the use of a device for producing a support material having chain molecules synthesized in synthesis fields, especially oligonucleotides which are complementary to segments of carriers of genetic information. According to the invention, after a synthesis run for synthesizing chain molecules (21) in a sequence of synthesis fields (1 to 16), at least one additional synthesis run is carried out, during which chain elements (n4 to n15) of one base-type sequence are linked to the chain molecules (21) synthesized during the preceding synthesis run or during each preceding synthesis run and comprising other chain elements (N1 to N12; N9 to N17) of another base-type sequence. The delivery of chain elements (N1 to N17; n4 to n15) to the synthesis fields (1 to 16) in the synthesis runs corresponds to the predetermined succession of chain elements (N1 to N17; n4 to n15) in the chain molecules (21) to be synthesized. The groups of synthesis fields (27) assigned to synthesis regions (1 to 16) are during each synthesis cycle displaced by one synthesis field (1 to 16), always moving in the same direction. In this way it is possible efficiently to synthesize, for example, chain molecules (21) which are complementary to at least parts of segments (20) of carriers of genetic information, with groups of chain elements (N1 to N17; n4 to n15) so as to obtain different base-type sequences.

Description

The invention relates to a method for producing a carrier material with in Syn chain molecules synthesized with a predetermined sequence of Chain building blocks, in particular of at least parts of sections of Erbinforma complementary oligonucleotides, in which in one synthesis step in a Sequence of synthesis cycles in a through direction relative to at least one reaction cell positioned to the carrier material with partially overlapping synthesis areas and for one Chain building blocks of a basic type series provided by the reaction cell be passed so that in several synthesis cycles in repeated from synthesis areas Synthetic fields covered by a reaction cell have chain molecules that are identical in sections Sequences of chain building blocks can be synthesized.

The invention further relates to a device in particular for producing a Carrier material with chain molecules synthesized in synthetic fields with a predetermined sequence of chain building blocks, in particular of at least parts of Sections of genetic information carriers complementary oligonucleotides, with one carrier material holder to which the carrier material can be applied, with a storage unit, with little least one reaction cell connected to the supply unit, through which the sealing cell Place the or each reaction cell on the carrier material for a chain extension provided chain building blocks in contact with the carrier material in a synthesis area can be passed through, with a positioning unit with which the or each reaction cell in one Synthesis run in a sequence of synthesis cycles each by one shift relative to the carrier material with synthesis partially overlapping in synthesis fields areas can be positioned, and with a control unit for controlling the supply unit and the Positioning unit, so that in several synthesis cycles in repeated from synthesis areas Synthesis fields of a reaction cell covered chain molecules with identical sections Sequences of chain building blocks can be synthesized.

The invention further relates to the use of a device for producing a Carrier material with chain molecules synthesized in synthetic fields with a predetermined sequence of chain building blocks, in particular of at least parts of  Sections of genetic information carriers complementary oligonucleotides, in particular for Carrying out a method according to one of claims 1 to 4, with a carrier material holder to which the carrier material is applied, with a supply unit, with little least one reaction cell connected to the supply unit, through which the sealing cell Place the or each reaction cell on the carrier material for a chain extension provided chain building blocks in contact with the carrier material in a synthesis area be passed through, with a positioning unit with which the or each reaction cell in one Synthesis run in a sequence of synthesis cycles each by one shift relative to the carrier material with synthesis partially overlapping in synthesis fields areas is positioned, and with a control unit for controlling the predicate unit and the Positioning unit, so that in several synthesis cycles in repeated from synthesis areas Synthetic fields covered by a reaction cell have chain molecules that are identical in sections Sequences of chain building blocks can be synthesized.

Such a method and such a device are known from the article "Arrays of complementary oligonucleotides for analyzing the hybridization behavior of nucleic acids "by E.M. Southern, S.C. Case-Green, J.K. Elder et al., Pages 1368 to 1373, Nucleic Acid Research, 1994, Vol. 22, No. 8, known. In the previously known method and previously known device is a reaction cell with a diamond-shaped or circular Synthesis area in one synthesis cycle in a sequence of synthesis cycles in one Direction of passage relative to a carrier material with partially overlapping synthesis layer range added, with a reaction solution with one for a ket chain extension provided in contact with the carrier material through the Reaction cell is passed. Chain molecules with a predetermined sequence of chains building blocks, in particular of oligo complementary to sections of genetic information carriers nucleotides can be synthesized by using a reaction solution with each of the synthesis cycles a predetermined chain building block in the synthesis area passed over the carrier material is so that in Syn covered repeatedly by synthesis areas of a reaction cell chain molecules with the same sequence of chain building blocks be synthesized.

The chain molecules are preferably designed as oligonucleotides. In the present The oligonucleotides of the serve on the carrier material (as a so-called "scanning array") Analysis of the hybridization behavior of nucleic acids. With the previously known method  and the previously known device are a variety of sequence homologues complementary Oligonucleotides with different chain lengths can be synthesized.

In inhibiting the expression of a protein encoded by a target nucleic acid Interaction (hybridization) of relatively short, to the base sequence of the target nucleic acid complementary oligonucleotides with the target nucleic acid or a target nucleic acid Protein complex, the target nucleic acid in particular a single-stranded target Nucleic acid with secondary and tertiary structure, especially a "messenger RNA" (mRNA) (so-called "antisense technology"), exists due to the complicated secondary and tertiary structure of the mRNA the problem of inhibiting the translation of a speci fish mRNA to identify available binding sites for an oligonucleotide, via their Localization is difficult to make predictions. A carrier material on which the arrangement of oligonucleotides mentioned can be in particular in an assay can be used to determine whether or how strong a target nucleic acid, how for example, such an mRNA molecule is able to target one or more of the to hybridize the oligonucleotides located on the carrier material. In such an assay If possible, the hybridization conditions are chosen so that the secondary or Tertiary structure of the target nucleic acid is largely preserved and with the corresponding in vivo structure of the RNA matches. The strength of the hybridization can be used as a measure of the Accessibility of a specific region of the target nucleic acid by the oligonucleotides serve. A single, identified as such, for hybridization with the target nucleic acid enabled oligonucleotide can then be prepared by known methods and in the Antisense technology for medical-therapeutic or diagnostic purposes be used.

From WO 95/21265 is known for the identification of usable for inhibition Binding sites as so-called "antisense" molecules hybridizable to an mRNA Synthesize oligonucleotides on a solid support, the oligonucleotides each completely from a single defined type of chain building blocks, for example from Deoxyribonucleotide type, deoxyribophosphorothioate nucleotide type or 2'-O-methyl Ribonucleotide type (i.e. each consisting of chain building blocks of a single basic type series) are.  

The known methods have the disadvantage that only oligonucleotides are found with them the carrier can be synthesized and identified, which are completely made up of chain molecules only certain basic type series are constructed. Such oligonucleotides often have not the multiple properties desired in antisense technology, such as high binding affinity for the target nucleic acid and / or high resistance to endogenous nucleases associated with the ability to express the target protein on the Effectively inhibit target nucleic acid level.

It is also known that pharmaceutically particularly efficient oligonucleotides are those which combine several properties, for example a high binding affinity for the target Nucleic acid / or resistance to degradation by endonucleases associated with the Property of efficient cleavage of the target nucleic acid by the endogenous RNAse H. cause. It has been possible to combine a large number of such properties in oligonucleotides and unite the same molecule by cutting such an oligonucleotide in sections Chain components of a certain basic type series is built, each section for a certain property or function of the oligonucleotide is responsible. Such Oligonucleotides are often referred to as "chimeric" oligonucleotides. Chimeric oligonucleotides consist of chain components of at least two different basic type series, which in at least two successive or adjacent sections, often also in three or more such sections are synthesized (see for example S.T. Crooke et al., J. of Pharmacology and Experimental Therapeutics 277 (1996), pp. 923-937).

The invention has for its object a method and a device of the beginning to create the type mentioned, in which an efficient synthesis of a Arrangement of chain molecules with identical sequences of chain building blocks different basic type series, in particular oligonucleotides with a chimeric structure, however otherwise largely the same characteristic characteristics as Base recognition characteristics can be performed on oligonucleotides.

The invention is based in particular on the task of forming sections of Genetic information carriers designed at least partially complementary oligonucleotides Chain molecules to create a method and a device of the type mentioned in the introduction, wherein the oligonucleotides have a chimeric structure, which results in a high Binding affinity and / or high resistance to such oligonucleotides  acting natural degradation enzymes, combined with the ability to inhibit efficiently translation, or more generally expression, of a target nucleic acid.

These tasks are solved with a method of the type mentioned in that after a sequence of synthesis cycles in one synthesis run at least one further Synthesis run with chain building blocks compared to the previous synthesis passage of different basic type series is carried out at the or everyone another synthesis run in synthesis cycles of a synthesis run one in this Reaction cell used reaction cell in the same direction as in the previous synthesis run is positioned so that the associated synthesis section by section with one previously uncovered in this synthesis run Synthesis field overlaid in the chain molecules with one in the or each previous one Synthesis pass synthesized maximum chain length exist, and that on this Provided chain position chain building blocks of the chain molecule to be synthesized in the this type of synthesis used basic type series passed through the reaction cell become.

These objects are achieved in a device of the type mentioned in that In the supply unit, chain building blocks before at least two different basic type series are advised that with the control unit, the or each reaction cell in at least two Synthesis passages can be positioned in the same passage directions in such a way that Synthesis cycles of a synthesis run, the associated synthesis area in sections overlaid with a synthesis field previously uncovered in this synthesis run, in which Chain molecules with a synthesis in the or each previous round of synthesis tized maximum chain length and that controlled by the control unit at two successive synthesis runs of chain building blocks of different basic type series can be passed through the or each reaction cell.

As an alternative to such a device, the storage unit of chain building blocks of at least two different basic type series, a device according to the invention can be a Corresponding supply unit, each chain components of a single one Contains basic type series, and in which the storage unit is designed to be interchangeable, so that after with each synthesis run, the storage unit with another storage unit Chain blocks of another basic type series can be exchanged.  

These objectives are achieved when using a device of the type described solved that in the supply unit chain building blocks at least two different reasons type series are stocked with the or each reaction cell with the control unit at least two synthesis runs in the same direction in each case so posi be tionation that during synthesis cycles of a synthesis run the associated Section in sections with a previously uncovered in this synthesis run Synthesis field overlaid in the chain molecules with one in the or each previous one Synthesis pass synthesized maximum chain length, and that from the tax Unit controlled chain building blocks in two successive synthesis runs different basic type series are passed through the or each reaction cell.

The present invention thus furthermore relates to the use of a device for producing a carrier material, comprising an arrangement of chain molecules synthesized in synthetic fields with a predetermined sequence of chain building blocks, in particular of oligonucleotides complementary to at least parts of sections ( 20 ) of genetic information carriers, with neighboring ones Synthesis fields include chain molecules with the same sequence of chain building blocks of different basic type series. Such an arrangement thus comprises chain molecules that comprise at least two sections, each with different basic type series of chain building blocks.

Another object of the invention is a carrier material, comprising an arrangement of chain molecules synthesized in synthetic fields with a predetermined sequence of chain building blocks, in particular oligonucleotides complementary to at least parts of sections ( 20 ) of genetic information carriers, with adjacent synthetic fields being chain molecules with the same sequence of chain building blocks of different basic type series include. Such an arrangement thus comprises chain molecules that comprise at least two sections, each with different basic type series of chain building blocks.

In a further aspect, the present invention is directed to a carrier material which can be produced by a method according to the present invention.  

By performing at least two superimposed synthesis runs according to the inventive method, the inventive device and the Use of a device according to the invention can be chain molecules with blocks of Chain blocks of different basic type series, but otherwise the same Synthesize characteristics such as base recognition characteristics, for example by different sequences of basic type series and / or different sizes designed synthesis areas used in the various synthesis runs Reaction cells high varieties in the synthesized chain molecules with high efficiency let achieve.

The synthesis areas of each reaction cell are expediently each rectangular equally large transverse sides, the length of each long side of a reaction cell corresponds to an integer multiple of a displacement path in the longitudinal direction. The Reaction cells are sized from one synthesis cycle to the next of a synthesis field offset in the longitudinal direction corresponding displacement. At The reaction cells are of different lengths for a first one Position the synthesis cycle of each synthesis run so that the synthesis areas of the Overlay the reaction cells flush with one another in the direction of passage.

In the context of the present invention, the chain molecules are, as already mentioned, preferably around oligonucleotides. Such oligonucleotides are for mating with one complementary strand of nucleic acid, especially an mRNA, and consist of individual chain building blocks, namely nucleosides or nucleoside analogues, which are usually a Nucleic acid base or a -base analog, which the properties of the oligonucleotides Convey base pairing, wear. A chain building block also includes an internucleosidic one Bridge binding to the next building block, for example from the phosphodiester, phosphorothioate or amide type, as described, for example, in A. de Mesmaeker et al., Acc. Chem. Res. 28 (1995), pp. 366-374.

Within the scope of the present invention there is a basic type of Chain molecules around chain building blocks of the same basic structure, with members of one first basic type series of members of a second basic type series, in particular through the Achieving properties, for example those of the part of the chain molecule under consideration  evoked or mediated physiological or pharmacological properties, differentiate. In the case of oligonucleotides, such properties are the chain molecules for example the affinity for a target nucleic acid, the nuclease resistance, the sensitivity versus RNAse H or the toxic properties. An advantageous oligonucleotide that acts as Drug used under antisense technology unites several beneficial properties in itself.

Within a chain molecule, such different properties are often linked to the presence of building blocks of certain basic structures or basic types. Examples of various basic type series of chain building blocks of oligonucleotides are known to the person skilled in the art. For example, the basic types of building blocks are natural nucleosides, such as 2'-deoxyribonucleosides or ribonucleosides; or by 2'-O-alkyl-modified ribonucleosides, the 2'-O-alkyl group, which preferably represents 2'-OC ₁ -C ₅ alkyl, in particular 2'-O-methyl or 2'-O-ethyl, unsubstituted or, in the case of 2'-O-ethyl at the β-position, can be substituted with -OH, -F or alkoxy, preferably with C ₁ -C ₅ alkoxy, in particular methoxy, where 2'-O- ( 2-methoxyethyl) modified ribonucleosides are very particularly preferred; around building blocks of nucleotides or nucleotide analogs which lead to oligonucleotides with an internucleosidic bridge binding of the phosphodiester or phosphorothioate type, or to oligonucleotides with a modified backbone (internucleosidic bridge binding), such as PNA building blocks, amide building blocks or 3'-methylenephosphonate Building blocks; or base-modified nucleoside building blocks, such as 5-propynyl nucleoside building blocks, the individual basic types having corresponding nucleic acid bases or base analogs depending on the base sequence of the target nucleic acid (see also A. De Mesmaeker et al., as mentioned above E. Uhlmann et al., Chem. Rev. 90 (1990), pp. 543-584). Combinations of chemical modifications lead to building blocks that belong to different basic type series. For example, 2'-O- (2-methoxyethyl) ribonucleotides with a phosphodiester group as building blocks of a basic type series, and 2'-O- (2-methoxyethyl) ribonucleotides with a phosphorothioate group as building blocks of another basic type series are considered in the context of the present invention. According to the invention, building blocks of a basic type series are synthesized in one section in an oligonucleotide, an oligonucleotide consisting of at least 2, for example 2 or 3 sections.

Chimeric oligonucleotides which consist of two sections are preferred as chain molecules, one section consisting of 2'-O- (2-methoxyethyl) ribonucleosides, which via Phosphodiester bridges or phosphorothloat bridges are linked together, and the other section consists of 2'-deoxyribonucleosides that cross over phosphorothioate bridges are linked together.

Chimeric oligonucleotides consisting of three sections are further preferred as chain molecules consist, whereby, viewed from 5'- in 3'-direction, the first section consists of 2'-O- (2- Methoxyethyl) -Ribonucleosiden exists that over phosphodiester bridges or Phosphorothioate bridges are linked together, the subsequent second Section consists of 2'-deoxyribonucleosides that bridge phosphorothioate with each other are linked, and the subsequent third section in turn consists of 2'-O- (2- Methoxyethyl) -Ribonucleosiden exists that over phosphodiester bridges or Phosphorothioate bridges are linked together.

In such chimeric oligonucleotides, the sections with 2'-O- (2-methoxyethyl- Containing ribonucleosides, two individual sections are preferred over a phosphorothioate Bridge is linked if the nucleoside is at the 3 'end of a section is a 2'-deoxyribonucleoside; or two separate sections are alternatively over one Phosphodiester bridge or linked together via a phosphorothioate bridge if it is the nucleoside at the 3 'end of a section is a 2'-O- (2-methoxyethyl) - Ribonucleoside.

Suitable, preferred chimeric oligonucleotides have a total length of about 15 to about 25, for example 20, nucleoside building blocks, the one consisting of 2'-deoxyribonucleoside building blocks existing section comprises at least about 5 such building blocks.

Further expedient refinements and advantages of the invention are the subject of Subclaims and the following description of exemplary embodiments below Reference to the figures of the drawing. Show it:

Fig. 1 is a schematic representation, in one embodiment a strip of carrier material with an attached, a four synthesis fields cover the synthesis area having reaction cell after a first Syn thesis cycle of a first synthesis passage with a graphical representation of the synthesized chain building blocks of a basic type series,

Fig. 2 shows the arrangement of FIG. 1 after performing the last synthesis cycle of the first synthesis passage,

Fig. 3 shows the arrangement according to FIG. 2 after performing a first synthetic cycle of another second synthesis passage with a concatenation Kettenbau a stone over or in the first synthesis through use different basic series,

Fig. 4 shows the arrangement according to FIG. 3 after performing the last synthesis cycle of the second synthesis passage,

Fig. 5 is a schematic representation in a further embodiment of a carrier strip of material with a a three synthesis fields covering Syn thesis area having reaction cell after performing a tenth synthesis cycle of a first synthesis passage showing the hitherto concatenated chain building blocks of a basic type series,

FIG. 6 shows the arrangement according to FIG. 5 with a reaction cell placed on top of it, which has a synthesis area covering five synthesis fields, after a second synthesis cycle of a further second synthesis run, FIG.

FIG. 7 shows the arrangement according to FIG. 6 after the last synthesis cycle of the second synthesis run has been carried out and

FIG. 8 shows the arrangement according to FIG. 7 with a reaction cell having a synthesis area covering two synthesis fields after the last synthesis cycle of a further third synthesis run has been carried out.

Fig. 9 shows a device for manufacturing a support material with synthesized in Synthesis fields chain molecules according to the invention in a schematic perspective view.

Fig. 10 shows the lifting unit of the device schematically in a perspective enlarged view.

Fig. 11, the lifting unit according to Fig 10 in a plan view.

Fig. A cross-section through the carrier bar of the lifting unit having a support beam attached to the reaction cell in a side view, partially cut away 12:

Fig. 13 is a plan view of a reaction cell

Fig. 1 of the invention in a schematic representation, in one embodiment, to a method and an apparatus according to a total of 16 Synthesis fields 1 assignable to 16 carrier strip of material 17 made, for example, chemical synthesis of oligonucleotides treated polypropylene (PP) as support material. The carrier material strip 17 is placed on a carrier material holder, not shown in FIG. 1, for example in the form of a metal plate provided with an elastic layer. Material strips onto the support 17 is a shown in outline displaceable reaction cell 18 placed in the representation according to Fig. 1 which is connected to an unillustrated in Fig. 1 stock unit. In further developments, a plurality of reaction cells 18 are provided which, for example for carrying out a parallel synthesis, are arranged next to one another on a reaction cell carrier and can be moved together. In the storage unit connected to a control unit (also not shown in FIG. 1), reaction fluids, for example reaction solutions, which are provided for a synthesis of chain molecules with chain extension with a predetermined, specific sequence of chain components, are stored or are successively stored, which are controlled by the control unit or can be fed to each reaction cell 18 .

The reaction cell 18 can be placed on the carrier material strips 17 with a seal. In the exemplary embodiment according to FIG. 1, the reaction cell 18 has a depression with an open-sided, rectangular, bordering a synthesis region 19 , the dimensions of which in the illustrated exemplary embodiment correspond to the size of four neighboring synthetic fields 1 to 16 in the illustration according to Fig. 1 is shown by means of a positioning unit, not shown in Fig. 1, controlled by the control unit in the direction of synthesis fields 1 to 16, with the reaction cell 18 being displaceable while being lifted, covering the first four synthesis fields 1 to 4 counted from the left.

In the following it is explained how a number of oligonucleotides complementary to at least part of a section 20 of an hereditary formation carrier are synthesized with the above-explained device in a method. The individual bases of section 20 carrying the genetic information are symbolically identified in FIG. 1 with B1 to B17 and each stand for one of the bases A, C, G and U or T according to the Watson-Crick model.

Further, 1 the structure is represented by a respective field synthesis 1 to 16 associated with oligonucleotides 21 and chain molecules in the representation of FIG.. Each oligonucleotide 21 is attached to a residual group R of the substrate strip 17 prepared for binding.

In the illustration of FIG. 1, the reaction cell 18 is disposed a first synthesis passage in the position of a first synthesis cycle, has been wherein an intended for a chain extension reaction fluid with a natural, complementary to the base B1 of the portion 20 nucleotide N1 through the reaction cell 18 is. The reaction fluid has washed over the synthesis area 19 , so that the nucleotide N1 has been added to the residual groups R of the first four synthesis fields 1 to 4 on the left in the illustration in FIG. 1.

1, the further synthesis cycles of the first synthesis run following the first synthesis cycle according to FIG. 1 are now, with the stepwise displacement of the reaction cell 18 by means of the positioning unit in a passage direction, in each case by a displacement path corresponding to the size of a neighboring synthesis field 5 to 16, the synthesis fields covered by the synthesis area 19 1 from left to right in the illustration according to FIG. 1 with natural reaction fluids containing nucleotides N2 to N13 of a basic type series complementary to the bases B1 to B13 of section 20 .

FIG. 2 shows the arrangement according to FIG. 1 with the reaction cell 18 in an arrangement that overlaps the four synthesis fields 13 to 16 on the left in the selected direction in the direction of passage after the last synthesis cycle of the first synthesis passage has been carried out with the one to the base B13 complementary natural nucleotide N13 freezing reaction fluid. From FIG. 2 it can be seen that, apart from the first three synthesis fields 1 to 3 and the last three synthesis fields 14 to 16 counted from the left in the representation according to FIG. 2, 4 to 13 sequence homologs in the central synthesis fields, each four natural nucleotides N1 to N13 having oligonucleotides have been synthesized 21, each to 13 synthesized oligonucleotide 21 as represented by the respective same sequence of ordinal numbers into a contiguous part of the section of the bases B1 is in a synthesis fields 4 20 to B13 of the genetic information carrier complementary.

FIG. 3 shows the arrangement according to FIG. 2 with the reaction cell 18 having the synthesis area 19 covering the four synthesis fields 1 to 16 in the positioning of a first synthesis cycle of a further second synthesis run. In the first synthesis cycle of the second synthesis run, the reaction cell 18 is brought into the same position as in the first synthesis cycle of the first synthesis run, covering the first four synthesis fields 1 to 4 on the left in the illustration in FIG. 3. A reaction fluid with a now modified nucleotide n5 as a chain building block of a different basic type series compared to the one used in the first synthesis run is then flushed out, the base of the modified nucleotide n5 being complementary to the base B5 of section 20 of the genetic information carrier. In the first synthesis cycle of the second synthesis run, an oligonucleotide 21 composed of five chain building blocks in the sequence N1-N2-N3-N4-n5 is synthesized in the fourth synthesis field 4, which is counted from the left, and which forms the first five bases B1 to B5 of the section 20 of the genetic information carrier is complementary and has natural nucleotides N1 to N4 of the basic type series used in the first synthesis run and the modified nucleotide n5 of the further basic type series used in the second synthesis run. In the edge-side synthesis fields 1 to 3 compared to the beginning of section 20 of the genetic information carrier not or not completely complementary oligonucleotides 21 have been synthesized.

In the further synthesis cycles of the second synthesis run following the first synthesis cycle, reaction fluids with modified nucleotides n6 to n17 are then passed through after shifting the reaction cell 18 each by a displacement path corresponding to the size of an adjacent synthesis field 5 to 16, where in the sequence of the bases B6 to B17 of section 20 of the genetic information carrier the sequence of the modified nucleotides n6 to n17 complementary to the bases B6 to B17 is predetermined.

FIG. 4 shows in the arrangement according to FIG. 3 the reaction cell 18 in the position in the last synthesis cycle of the second synthesis run after flushing with a reaction fluid containing the modified nucleotide n17 complementary to the last base B17 of section 20 of the genetic information carrier. From Fig. 4 it can be seen that now except for the three edge-side synthesis fields 1, 2, 3, 14, 15, 16 in the middle synthesis fields 4 to 13 sequence homologues, each with a part of section 20 of the genetic information carrier complementary oligonucleotides 21 with a Length of eight nucleotides have been synthesized. Each oligonucleotide 21 has four natural nucleotides N1 to N13 in blocks and then four modified nucleotides n5 to n17, each following the natural nucleotides N4 to N13 added last in the first synthesis run in one of the sequences of bases B1 to B17 of section 20 of the Genetic information carrier corresponding order.

The carrier material strip is now, in particular with regard to the oligonucleotides 21 synthesized in the middle synthesis fields 4 to 13, as chain molecules with eight chain units arranged in two blocks or sections with different basic type series, for use in an assay for examining parts of section 20 of the genetic information carrier with regard to accessible binding sites for so-called "antisense" oligonucleotides, as mentioned above.

Fig. 5 of the invention shows schematically similar to Fig. 1 to Fig. 4 in another embodiment, to a method and apparatus in accordance with the carrier strip of material 17 according to the aforementioned embodiment. In the above-referred embodiment will turn to connected parts of the portion 20 of the genetic information carrier complementary oligonucleotides 21 with a relation to the explained with reference to Fig. 1 to Fig. 4 embodiment greater chain length using natural and modified nucleotides, in three different long blocks of pairwise different basic type series be synthesized.

For this purpose, as shown in FIG. 5, a reaction cell 22 is shown with a three-synthesis fields 1 to 16 covering synthesis portion 23 in the array in a tenth cycle of the synthesis of a first synthesis passage. In the first synthesis cycle of the first synthesis run, the reaction cell 22 was arranged in such a way that the third to fifth synthesis fields 3, 4, 5 counted from the left in the illustration according to FIG. 5 was covered by the synthesis region 23 . Then, in the subsequent synthesis cycles of the first synthesis run, the reaction cell 22 was placed in each case on the next neighboring synthesis field 5 to 14 and flushed with a reaction fluid containing a complementary natural nucleotide N2 to N10 corresponding to the sequence of bases B2 to B10, so that up to the in tenth synthesis cycle shown Fig. 5, the comple mentary oligonucleotides 21 shown have built up.

FIG. 6 shows the arrangement according to FIG. 5 after a second synthesis cycle of a further second synthesis run. In the second synthesis step, a reaction cell 24 is used, with the synthesis area 25 of which five synthesis fields 1 to 16 can be covered. In a first synthesis cycle of the second synthesis run, the reaction cell 24 is arranged such that the only synthesis field 5 to 12 with a chain of three natural nucleotides N1 to N12 is the fifth synthesis field 5 counted from the left in the illustration in FIG. 6 together with the left side outer four synthesis fields 1 to 4 has been covered for the first time by the synthesis area 25 . In the first synthesis cycle of the second synthesis run, the first two synthesis fields 1, 2 located on the left outside in the illustration in FIG. 6 were first flushed with a reaction fluid.

In the first synthesis cycle of the second synthesis run, a reaction fluid was used which contains the modified nucleotide n4 which is complementary to the fourth base B4 of section 20 , while in the second synthesis cycle of the second synthesis run shown in FIG. 6 that to the fifth base B5 of section 20 of the genetic information carrier complementary, modified nucleotide n5 was contained in the reaction fluid.

In the further synthesis cycles of the second synthesis run, the reaction cell 24 is in each case displaced by a displacement path corresponding to the size of a synthesis field 1 to 16, and reaction fluids with complementary modified nucleotides n6 to n15 in the corresponding sequence of the bases B6 to B15 of section 20 of the genetic information carrier are passed through .

Fig. 7 shows in the representation of Fig. 6 the positioning of the reaction cell 24 used in the second synthesis run after performing the last synthesis cycle of the second synthesis run. From Fig. 7 it can be seen that apart from the four synthesis fields 1 to 4, 13 to 16 located on the right and left edges in the middle synthesis fields 5 to 12 oligonucleotides with eight chain components N1 to N10, n4 to n15 have been synthesized, the in the first synthesis step, three nucleotides N1 to N10 synthesized in blocks from a natural basic type series and the five nucleotides n4 to n15 linked in the second synthesis process in blocks are assigned to a modified basic type series.

FIG. 8 shows the arrangement according to FIG. 7 after carrying out the last synthesis cycle of a further third synthesis run, in which, as chain building blocks, a basic type series different from the previous synthesis run in turn complements the natural nucleotides N9 to N17 to the bases B9 to B17 to those in the first two Synthesis runs synthesized oligonucleotides 21 have been linked. In the third synthesis step, a reaction cell 26 is used, with the synthesis region 27 of which two adjacent synthesis fields 1 to 16 can be covered.

In a first synthesis cycle of the third synthesis run, the reaction cell 26 was positioned such that the fifth synthesis field 5 counted from the left in the illustration in FIG. 8 was the only synthesis field covered for the first time with a full number of eight chain building blocks synthesized in the previous two synthesis runs . In this first synthesis cycle, a reaction fluid with a natural nucleotide N9 complementary to the ninth base B9 of section 20 of the genetic information carrier was passed through the reaction cell 26 .

The further synthetic cycles of the third synthesis passage of the reaction cell 26 were field for the initial covering of the respective next adjacent Synthesis 6 to 16 by the sequence of bases B10 to B17 predetermined complementary natural nucleotides flushed N10 to N17 containing reaction fluids through the reaction cell 26 by displacement.

As can be seen from FIG. 8, 5 to 12 sequence homologous oligonucleotides 21 with a length of ten chain building blocks are now synthesized in the middle synthesis fields after completion of the third synthesis step, each of which is complementary to part of section 20 of the genetic information carrier and consists of a block of three natural nucleotides N1 to N10, a block of five modified nucleotides n4 to n15 and a further block of two natural nucleotides N9 to N17 are constructed. Each block of natural nucleotides N1 to N10 or N9 to N17 and of modified nucleotides n4 to n15 has a length which corresponds to the number of synthesis fields covered by the synthesis areas 23 , 25 , 27 of the reaction cells 22 , 24 , 26 used in the corresponding synthesis runs Corresponds to 1 to 16.

It goes without saying that in further, not shown versions longer chain Chain molecules by using larger synthesis areas Reaction cells using a larger number of synthesis cycles more synthesis runs than those detailed above Embodiments can be synthesized. By providing reaction cells with differently sized synthesis areas are diverse variations in the block Sequence of basic type series can be achieved, it being expedient that at rectangular trained synthesis areas the transverse sides of the reaction cells and the same size Size of the long sides a full-page multiple of that by a displacement in Direction of passage specified dimension of the synthesis fields correspond and the Reak tion cells offset in the longitudinal direction by the corresponding dimension of the synthesis fields become.

It is understood that more than two different basic type series in the different Synthesis runs can be provided so that with an appropriate number the most varied variations in the sequence of basic type series are cash.  

Examples 1. Production of an activated polypropylene carrier material

A cut polypropylene film (PP: Mobil 40MB400, 400 mm × 300 mm) is placed in a cylindrical glass drum (diameter: 170 mm, length: 400 mm) in a solution (180 ml) of 0.9 g chromium (VI) oxide (Fluka), dissolved in a 1: 1 mixture of acetic anhydride: acetic acid (Fluka), immersed. The reaction drum is quickly rotated for 1 hour using a mechanical rolling device. The film is then removed, immersed in a bath of 1 M aqueous acetic acid for 30 minutes and then rinsed with methanol (Fluka) for 30 minutes. The film is dried in a high vacuum, being located between 2 glass plates for protection. The film is then immersed in a corresponding glass container in a 1 M solution of BH ₃ (Aldrich Chemical Co.) in tetrahydrofuran (180 ml), which is diluted to 0.2 M with additional tetrahydrofuran. The glass jar is rotated quickly for 1 hour. The film is removed, immersed in 1 M aqueous acetic acid, rinsed in methanol and dried under vacuum. The film treated in this way is used without further treatment as a carrier material in a process according to the invention.

2. Automated device for producing a carrier material according to the invention with in Synthetic fields synthesized chain molecules

In Fig. 9 an automated apparatus (so-called. "Array Maker") is shown, which is used for the synthesis of an inventive support material with synthesized in Synthesis fields chain molecules. The device comprises an automatic cell positioning device or cell positioning unit 36 and a DNA synthesizer (ABI 394/4) 31. The DNA synthesizer 31 is connected to the automatic cell positioning device 36 by means of hose connections for the reagents (not particularly reaction solutions) which are not shown in the drawing , Purge liquids and purge gases). The inlet tubing connections that are normally used to deliver the reagents from the DNA synthesizer to the 4 reaction columns, which comprise a solid support and are standard for the synthesis of oligonucleotides, are instead with the inlet openings of 4 reaction cells 32 , 33 , 34 , 35 connected. The outlet tube connections, which normally connect the DNA synthesizer to the respective outlet openings of the 4 reaction columns mentioned, are instead connected to the outlet openings of the 4 reaction cells 32 , 33 , 34 , 35 . The cell positioning machine 36 is connected via a trigger line (not shown in the drawing) to an electronic control unit 37 , with the aid of which a trigger signal ("advance fraction collector signal") for the next cell position of the cell positioning machine 36 from the DNA synthesizer 31 the cell positioning machine 36 is transmitted. The automatic cell positioning device 36 and the DNA synthesizer 31 stand on a stable base 38 below a hood 39 with a rear wall 40 . The automatic cell positioning device 36 has its own protective hood 41 , which is only shown in parts in the drawing, and which is provided with a vertical sliding door, which is operated with the aid of a handle 42 and via pulling ropes, not shown in the drawing, with one in the drawing Counterweight, not shown, are connected and can be deflected via rollers 43 , can be moved into an indicated upper and also an indicated lower position. The rollers 43 are connected via brackets and struts to the profile frame 45 consisting of several profile tubes 44 and supported on the base 38 . The automatic cell positioning device ( 36 ) is moved by an electrically operated unit 46 , which, controlled by the control unit 37 , moves a pressure plate 47 vertically in stages between an illustrated upper position and an indicated lower position.

The PP film 49 is fastened on the pressure plate 47 with the aid of 5 vertical metal holding bars 48 by means of screws located at the upper and lower ends of the holding bars. A thin piece of rubber 90 (see FIG. 11) is located between the PP film 49 and the pressure plate 47 . The pressure plate 47 is moved vertically to preprogrammed positions. The reaction cells 32 , 33 , 34 and 35 are mounted horizontally next to one another on a support bar 50 of a lifting unit 51 .

The lifting unit 51 is shown separately and enlarged in FIG. 10. Fig. 11 shows the lifting unit 51 in plan view together with the reaction cells 32, 33, 34 and 35 and the pressure plate with the 47th The lifting unit 51 has a left guide 52 and a right guide 53 , which are each designed as a hollow cylinder. A left push rod 54 and a right push rod 55 each extend through the guides 52 and 53 . Both pressure rods 54 , 55 are biased with the aid of pressure springs (not shown in the drawing) arranged in the guides 52 , 53 in such a way that the reaction cells 32 , 33 , 34 , 36 are pressed onto the pressure plate 47 . To lift the reaction cells 32 , 33 , 34 , 35 away from the pressure plate 47 , eccentric disks, not shown in the drawing, are provided which cooperate with the end faces 56 and 57 of the pressure rods 54 , 55 , around the support bar 50 and thus the reaction cells 32 , 33 , 34 , 35 to move away from the pressure plate 47 . The eccentric discs are adjusted with the aid of an electric drive, which is also connected to the control electronics 37 .

The push rods 53 , 54 are detachably connected to the support beam 50 by means of knurled nuts 58 . In this way, the reaction cells 32 , 33 , 34 , 35 can be removed together with the support bar 50 . After the support bar 50 has been removed, it is possible to detach each individual reaction cell 32 , 33 , 34 , 35 from the support bar 50 and, if appropriate, to replace it with another reaction cell 32 , 33 , 34 , 35 .

The connection of the reaction cell 32 is shown in FIG. 12, for example. The reaction cell 32 is fastened to a pin 59 which extends through a bore 60 in the support beam 50 . On the side of the support beam 50 pointing away from the reaction cell 32 , the pin 59 is provided with a thread onto which a knurled nut 61 is screwed. The pin 59 with the screwed-on knurled nut 61 is prestressed with the aid of a compression spring 62 , so that the knurled nut 61 is pressed against the back of the support beam 50 . The compression spring 62 is designed to be considerably weaker than the compression springs provided in the guides 52 , 53 , so that when the reaction cells 32 , 33 , 34 , 35 are placed on the plastic film 49 and the pressure plate 47, the compression spring 52 is compressed and the pressure force of the reaction cell 32 , 33 , 34 , 35 . The optimal pressure force can be set quickly and easily by changing the pressure spring 62 . A compression spring 62 with 42N is used for the PP film 40MB400 used as carrier material in this example. In addition, the compression spring 62 ensures a uniform pressure of the reaction cells 32 , 33 , 34 , 35 against the plastic film 49 (see FIG. 11), which either directly on the pressure plate 47 or with the interposition of an intermediate carrier film 90 (see FIG. 11) elastic material is fastened to the pressure plate 47 with the aid of the holding webs 48 . The intermediate carrier film 90, which is preferably up to 1 mm thick (see FIG. 11), consists, for example, of a polymer film made of Neoprene 60 ° Shore A or natural rubber (for example Paragummi 40 ° Shore A), both from Richterich + Zeller AG, Basel, Switzerland are distributed.

The structure of the reaction cells 32 , 33 , 34 , 35 , which are open on one side, is explained in more detail below with reference to FIGS. 12 and 13.

As can be seen in FIGS. 12 and 13, the reaction cells 32 , 33 , 34 , 35 consist of a block with a rectangular cross section made of a deformable material or a plastic. In this case, PTFE is used. The rear side 63 facing the support beam 50 and the outer sides 64 , 65 , 66 , 67 and 68 are each flat. On the upper side 69 pointing away from the support beam 50 , a peripheral edge strip 70 is provided, which encloses a flat space 71 (corresponding to 19 , 23 , 25 , 27 in FIGS. 1 to 8). The height of this flat space 71 and the height of the edge strip 70 are one millimeter. The four edge strip sections circumscribing a rectangle are each 40 mm × 10 mm in size. In the area of the corners 72 , 73 , 74 and 75 , the edge strip sections run in an arc shape, the inner radius being approximately 0.5 millimeters.

Between the corners 72 and 75 or 73 and 74 there are bores 76 and 77 running at right angles to the upper side, which in the exemplary embodiment shown likewise have a radius of 0.5 millimeters. The bore 76 opens into an outlet channel 78 . The lower bore 77 opens into an inlet channel 79 . Each inlet channel 79 and each outlet channel 78 of the reaction cells 32 , 33 , 34 , 35 is connected to the synthesizer 1 via a hose connection, not shown in the drawing.

3. Operation of the synthesis device

During operation of the device, the support bar 50 is moved horizontally in order to press the reaction cells 32-35 against the PP film. When an electrical signal is received from the advance fraction collector, the control unit 37 sends a signal to the cell positioning machine 36 . The support bar 50 is then moved horizontally to remove the reaction cells 32-35 from the surface of the PP film. The pressure plate 47 is then moved vertically into a new position. Finally, the carrier bar 50 is again moved horizontally, and the reaction cells 32-35 are pressed again onto the surface of the PP film.

The preparation of an arrangement of chimeric oligonucleotides on the solid support material comprises the following steps:

• 1. A PP film 49 preactivated as in Example 1 is fastened in the cell positioning machine 36 using a rubber piece 90 in order to guarantee a good seal.
• 2. The vertical support bars 48 are attached in place to ensure that the film 49 holds well.
• 3. The selected reaction cells (between 1 and 4) 32 , 33 , 34 , 35 , with corresponding compression springs 62 are attached to the support beam 50 , which is installed on the cell positioning machine.
• 4. The selected reaction cells 32 , 33 , 34 , 35 are connected to the inlets and outlets of the DNA synthesizer via hose connections.
• 5. The coordinates of the starting position and the length of the displacement path are programmed into the control unit 37 , the pressure plate 47 is moved into the starting position, and the reaction cells 32 , 33 , 34 , 35 are brought into contact with the PP film 49 .
• 6. The DNA synthesizer is programmed so that the desired ones Oligonucleotide sequences can be synthesized in the desired arrangement, and the Synthesis begins.
• 7. As soon as the so-called "advance fraction collector" signal is generated according to the synthesis protocol (see below), the lifting unit 51 lifts the reaction cells away from the surface of the PP film, the pressure plate moves vertically upwards by the selected displacement length, and moves the lifting unit 51 forwards the support beam 50 , whereby the contact between the reaction cells and the film is established. A reaction cycle is then carried out by means of the DNA synthesizer in order to synthesize a first building block of an oligonucleotide on the surface of the film or in order to carry out a chain extension reaction of an oligonucleotide.
• 8. Step 7 is repeated as many times as necessary for a synthesis run.

4. Production of an arrangement of oligonucleotides on a carrier (so-called "scanning array"). which comprises chimeric oligonucleotides with a length of 16 nucleotide building blocks

The PP film 49 is pretreated as described herein. A sheet of natural rubber (Paragummi 40 ° Shore, Richterich & Zeller, Basel, Switzerland, 0.5 mm thick, 400 × 300 mm) 90 is used as described. As described above, 4 rectangular reaction cells 32 , 33 , 34 , 35 made of Teflon are used, each reaction cell being held by two compression springs 62 ( 42 N). Each reaction cell is connected to the inlet and outlet tubing connections of an ABI 394/4 DNA synthesizer (Perkin Elmer Corp., Foster City, CA, USA). The starting position of the beam is set to 5 mm below the top edge of the PP film. 5 mm are programmed as the length of the displacement path of the reaction cells after each synthesis cycle. Due to the length of the reaction cells of 40 mm, an arrangement of oligonucleotides is generated in one synthesis run, which have a maximum length of 8 building blocks.

The reaction sequences described below result in an arrangement of chimeric oligonucleotides (total length of a chimeric oligonucleotide: 16 nucleotide building blocks, of which (i) building blocks 1 to 8: deoxyribonucleosides, which are linked to one another via phosphorothioate bridges, and (ii) building blocks 9 to 16 :
2'-O- (2-methoxyethyl) ribonucleosides which are linked to one another via phosphodiester bridges). In analogy to the procedure described in general form above ( FIGS. 1 to 3; chimeric oligonucleotide with a total length of 8 building blocks, of which the first 4 are called N1 to N4 and the last 4 are called N5 to n8), the nucleotides of the in a part of the desired oligonucleotides comprising nucleotides N1 to N8 produced in a first synthesis step and comprising blocks 1 to 8, while the nucleotides of the part of the desired oligonucleotides comprising blocks 9 to 16 produced in the second synthesis step would be referred to analogously as n10 to n16.

The storage unit includes 8 bottles with nucleoside building blocks, 4 of which Bottles one of the four deoxyribonucleotides (corresponds to chain building blocks of a first Basic type series) included, and the 4 remaining bottles each one of the four 2'-O- (2- Methoxyethyl) ribonucleosides (corresponds to chain building blocks of a second basic type series) contain. In the following, the synthesis of the 1st to 8th building blocks of the Oligonucleotides the corresponding bottles with the building blocks of the first basic type series controlled, and for the synthesis of the 9th to 16th building blocks, the corresponding Driven bottles with the components of the second basic type series.

• (i) Synthesis of the 1st to 8th building blocks of an arrangement of oligonucleotides on the PP film (DNA building blocks linked via phosphorothioate bridges)
  The synthesis of the DNA phosphorothioate portion of the oligonucleotides is accomplished by programming the DNA synthesizer to use one of the four possible reaction cells, for example cell 32 , to generate the desired arrangement of oligonucleotides. The three remaining reaction cells, for example 33 , 34 and 35, are freely available in order to simultaneously synthesize adjacent arrays of oligonucleotides on the same PP film which correspond to other target sequences. Commercially available phosphoramidites are used as nucleotide building blocks (Cruachem, A: 2081120-21; G: 208199-2; C: 208130-21; T: 208100-21). The synthesis machine dilutes 2 g of the phosphoramidites with 50 ml of acetonitrile (MWG-Biotech). In the synthesis, tetrazole (Cruachem. 17112044) as used by the manufacturer is used; Trichloroacetic acid in 1,1,1-trichloroethane (2.5% w / v) is used for detritylation; Phenylacetyl disulfide (prepared according to p. 329 of the article by HCPF Roclen et al., Recl. Trav. Chim. Pays-Bas 110 (1991), p. 325-331) is used as a 0.5 M solution in a mixture of acetonitrile / 3 -Picolin (4: 1) used for oxidation. The first base (from the 3 'end) is coupled to the PP film at the start position, and subsequent bases are added by the reaction cell moving vertically downwards in accordance with the correspondingly programmed control unit.

An arrangement of oligonucleotides is generated which is based on the following sequence:
5'-GGC CTG AAG GTG CTG GAC AGT GTC TGA AAG TAG GGC ACT AGC CAA GTT GGA GCA GAA GGC-3 '(SEQ ID NO 1).

In the form shown, this sequence is complementary to the sequence of the target nucleic acid. This sequence is programmed into the synthesizer, the synthesis of 3'- in 5'- Direction.

The synthesis protocol reproduced below for coupling a nucleotide building block to the film or to a nucleotide chain (ie a synthesis cycle) is a protocol of the DNA synthesizer ABI 394/4, which is based on the specific circumstances of the production of an arrangement of oligonucleotides has been adapted on a PP film. The protocol is reproduced in English, since the machine must be programmed in English:

Bottle 18 contains acteonitrile, bottle 14 contains the detritylation solution, bottle 10 contains the phenylacetyl disulfide reagent and bottle B contains the respective phosphoramidite solution.

The protocol described is used correspondingly often to couple the other modules repeated until the desired arrangement of the first part of chimeric oligonucleotides is completed, the first part, based on a single oligonucleotide, at most each comprises the first 8 building blocks, and the overall arrangement is as given above Sequence includes.

• (ii) Synthesis of the 9th to 16th building blocks of an arrangement of oligonucleotides on the PP film (2'-O- (2-methoxyethyl) ribonucleosides linked via phosphodiester bridges)
  The reaction cell is reset to the original starting position. The DNA synthesizer is programmed so that 2'-O- (2-methoxyethyl) ribonucleoside-phosphoramidite building blocks (2 g) are used for the 9th to 16th building blocks, which according to P. Martin, Helv. Chim. Acta 78 (1995), pp. 486-504 and dissolved in 50 ml of acetonitrile. The tetrazole (Cruachem. No. 17112044) used in the further course of the synthesis is used as obtained from the manufacturer. Trichloroacetic acid in 1.1.1 trichloroethane (2.5% w / v) is used for demetilation. A mixture of iodine / lutidine / water / acetonitrile is used for the oxidation (0.65 g (Fluka) / 50 ml (Fluka) / 2.5 ml / 450 ml). The synthesis protocol is modified according to the use of the modified phosphoramidite building blocks. The first base (3 'end) in each sequence is coupled to the PP film at the starting position of the reaction cell. The starting position corresponds to the starting position at the beginning of the first synthesis run (5 mm below the top edge of the PP film). Subsequent bases are added while the reaction cell is moved vertically along the carrier film (displacement length 5 mm). The synthesis sequence is based on the following base sequence:
  5'-ccc acc gag gcc tga agg tgc tgg aca gtg tct gaa agt agg gca cta gcc aag ttg cag-3 '(SEQ ID NO 2).

This sequence is programmed into the synthesizer; the synthesis takes place again in the 3'-5 'direction. The first base 'g' at the 3 'end corresponds to the SEQ ID No 2 of the 9th base "G" counted from the 3 'end of the SEO ID NO 1, since the maximum length of the im 1. Synthesis pass synthesized oligonucleotides is 8 chain building blocks.

Synthesis protocol of the synthesis machine ABI 394/4 (modified with regard to the production of the oligonucleotide arrangement ("array"); the "advance fraction collector" signal is used to shift the reaction cell). The protocol is reproduced in English, since the machine must be programmed in English:

Bottle 18 contains acteonitrile, bottle 14 the detriylation solution, bottle 15 the oxidation reagent (I ₂ / lutidine / H ₂ O) and bottle B the respective phosphoramidite solution.

In this way, the desired arrangement is finally obtained ("scanning array") of chimeric oligonucleotides on the support.

5. Deprotection of the oligonucleotides

The PP film is removed from the device and rinsed in isopropanol. After that the film is placed in 32% ammonia at room temperature in a reaction drum (see Example 1) washed. The chamber is rotated quickly for 16 hours to get a good one Guarantee mixing of the reagents. The film is removed from the chamber with Rinsed water and methanol and dried briefly under vacuum. The slide with the desired arrangement of chimeric oligonucleotides can now be in a standard Hybridization assay, as mentioned above, can be used.

Claims (14)

1. A method for producing a carrier material with chain molecules synthesized in synthesis fields with a predetermined sequence of chain building blocks, in particular oligonucleotides complementary to at least parts of sections ( 20 ) of genetic information carriers, in which at least one in a synthesis cycle in a sequence of synthesis cycles in one cycle direction Reaction cell positioned relative to the carrier material ( 17 ) with partially overlapping synthesis areas and chain building blocks of a basic type series intended for chain extension are passed through the reaction cell, so that chain molecules with the same sequence of chain building blocks are synthesized in several synthesis cycles in synthesis fields repeatedly covered by synthesis areas of a reaction cell, characterized in that after a sequence of synthesis cycles in one synthesis run, at least one further synthesis run with Ke It is carried out in a basic type series which is different from the previous synthesis run, in such a way that in the or each further synthesis run during synthesis cycles of a synthesis run, a reaction cell ( 18 , 22 , 24 , 26 ) used in this synthesis run is positioned in the same pass direction as in the previous synthesis run is that the associated synthesis area ( 19 , 23 , 25 , 27 ) is overlaid in sections with a synthesis field (1 to 16) hitherto uncovered in this synthesis run, in which chain molecules ( 21 ) with a maximum chain length synthesized in the or each previous synthesis run are present , and that at this chain position provided chain building blocks of the chain molecule to be synthesized of the basic type series used in this synthesis run are passed through the reaction cell ( 18 , 22 , 24 , 26 ). 2. The method according to claim 1, characterized in that in a first synthesis cycle of the or each further synthesis pass, the or each reaction cell used in this synthesis pass ( 18 , 22 , 24 , 26 ) is positioned so that a single synthesis field (1 to 16 ) with chain molecules ( 21 ) of a maximum chain length synthesized in the or each synthesized prior to the beginning of the synthesis run of their synthesis region ( 19 , 23 , 25 , 27 ), and that with each further synthesis cycle a further adjacent synthesis field (3 to 16) with a maximum chain length synthesized in the or each synthesis step before the synthesis step is superimposed for the first time by the synthesis area ( 19 , 23 , 25 , 27 ). 3. The method according to claim 1 or 2, characterized in that in each synthesis step, the or each reaction cell ( 18 , 22 , 24 , 26 ) is displaced by the same displacement path in each synthesis cycle, so that the synthesis fields (1 to 16) are of the same size are. 4. The method according to claim 3, characterized in that in at least two synthesis runs, the synthesis areas ( 23 , 25 , 27 ) of the reaction cells used ( 22 , 24 , 26 ) are of different sizes, each synthesis area ( 23 , 25 , 27 ) in one direction corresponds to a multiple of the dimensions of the synthesis fields (1 to 16) in this direction. 5. Device in particular for producing a carrier material with chain molecules synthesized in synthetic fields with a predetermined sequence of chain building blocks, in particular oligonucleotides complementary to at least parts of sections ( 20 ) of genetic information carriers, in particular for carrying out a method according to one of claims 1 to 4, with a Carrier material holder to which the carrier material ( 17 ) can be applied, with a storage unit, with at least one reaction cell connected to the storage unit, through which, when the or each reaction cell is sealingly placed on the carrier material ( 17 ), chain modules provided for a chain extension in contact with the Carrier material ( 17 ) can be passed through in a synthesis area, with a positioning unit with which the or each reaction cell in a synthesis run in a sequence of synthesis cycles each by a displacement path relative to the carrier material erial ( 17 ) can be positioned with synthesis areas partially overlapping in synthesis fields, and with a control unit for controlling the storage unit and the positioning unit, so that in the case of several synthesis cycles in synthesis fields repeatedly covered by synthesis areas of a reaction cell, chain molecules with sections of identical sequences of chains can be synthesized, thereby characterized in that chain components (N1 to N17, n4 to n15) of at least two different basic type series are stored in the supply unit, so that the or each reaction cell ( 18 , 22 , 24 , 26 ) can be positioned with the control unit in at least two synthesis cycles in the same direction in each case is that in synthesis cycles of a synthesis pass, the associated synthesis area ( 19 , 23 , 25 , 27 ) is partially overlaid with a synthesis field (1 to 16) previously uncovered in this synthesis pass, in which chain molecules ( 21 ) with a are present in the or each previous synthesis passage, the maximum chain length synthesized, and that the control unit controls ge in two successive synthesis passes chain building blocks (N1 to N17, n4 to 15) of different basic type series through the or each reaction cell ( 18 , 22 , 24 , 26 ) are. 6. The device according to claim 5, characterized in that the or each reaction line ( 18 , 22 , 24 , 26 ) has a rectangular synthesis area ( 19 , 23 , 25 , 27 ). 7. The device according to claim 6, characterized in that the longitudinal dimensions of each synthesis area ( 19 , 23 , 25 , 27 ) of a reaction cell ( 18 , 22 , 24 , 26 ) an integer multiple of the size of the synthesis fields determined by the same displacement paths in all synthesis runs (1 to 16) in the longitudinal direction of the synthesis areas ( 19 , 23 , 25 , 27 ) corresponds. 8. Device according to one of claims 5 to 7, characterized in that reaction cells used in different synthesis runs ( 22 , 24 , 26 ) have differently sized synthesis areas ( 23 , 25 , 27 ). 9. Use of a device defined in claim 5 for the production of a carrier material comprising an arrangement of chain molecules synthesized in synthetic fields with a predetermined sequence of chain building blocks, in particular of at least parts of sections ( 20 ) of genetic information carriers complementary oligonucleotides, with neighboring synthetic fields containing chain molecules with sections same sequence of chain components of different basic type series. 10. Use of a device according to claim 9, characterized in that the or each reaction cell ( 18 , 22 , 24 , 26 ) has a rectangular synthesis area ( 19 , 23 , 25 , 27 ). 11. Use of a device according to claim 10, characterized in that the longitudinal dimensions of each synthesis area ( 19 , 23 , 25 , 27 ) of a reaction cell ( 18 , 22 , 24 , 26 ) an integer multiple of the size determined by the same displacement paths in all synthesis runs the synthesis fields (1 to 16) in the longitudinal direction of the synthesis areas ( 19 , 23 , 25 , 27 ) corresponds. 12. Use of a device according to one of claims 9 to 11, characterized in that reaction cells used in different synthesis runs ( 22 , 24 , 26 ) have differently sized synthesis areas ( 23 , 25 , 27 ). 13. Carrier material, comprising an arrangement of chains synthesized in synthesis fields with a predetermined sequence of chain building blocks, in particular of oligonucleotides complementary to at least parts of sections ( 20 ) of genetic information carriers, adjacent synthesis fields comprising chain molecules with the same sequence of chain building blocks of different basic type series. 14. A carrier material comprising an arrangement of chain molecules synthesized in synthetic fields with a predetermined sequence of chain building blocks, in particular of oligonucleotides complementary to at least parts of sections ( 20 ) of genetic information carriers, which can be produced by a method according to one of claims 1 to 4.