CN218077815U - Novel oligonucleotide synthesis system - Google Patents

Novel oligonucleotide synthesis system Download PDF

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CN218077815U
CN218077815U CN202221346930.9U CN202221346930U CN218077815U CN 218077815 U CN218077815 U CN 218077815U CN 202221346930 U CN202221346930 U CN 202221346930U CN 218077815 U CN218077815 U CN 218077815U
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pipeline
conveying pipeline
way valve
conveying
inlet
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周胜
徐慧
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Lisure Science Suzhou Co ltd
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Lisure Science Suzhou Co ltd
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    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
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    • Y02P20/50Improvements relating to the production of bulk chemicals
    • Y02P20/55Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

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Abstract

The utility model discloses a novel oligonucleotide synthesis system. The system comprises a first pipeline, a second pipeline, a third pipeline, a circulating pipeline, a liquid inlet pipeline, a synthetic column, a liquid outlet pipeline and a bypass pipeline. The novel oligonucleic acid synthesis system can realize residue-free and residue-free cleaning when synthesizing DNA, and accurately control the volume through continuous constant current; and a synthesis column with variable volume is also arranged, so that large-volume circulating reaction is realized, and the reaction efficiency is improved.

Description

Novel oligonucleotide synthesis system
Technical Field
The utility model relates to an oligonucleotide synthesis field, in particular to a novel oligonucleotide synthesis system.
Background
The fragments synthesized by DNA are often expressed by different methods, such as oligonucleotides, PCR primers, linkers, probes, short-chain primers, long-chain primers, etc., and in the laboratory, DNA synthesis is generally performed by various equipment tools, such as a DNA synthesizer, a DNA synthesis column, etc.
The process of DNA synthesis is briefly described as follows:
in the first step, the nucleotide with protected active group previously attached to the solid phase carrier is reacted with trichloroacetic acid to remove the 5 '-hydroxyl protecting group DMT, and the free 5' -hydroxyl is obtained.
And secondly, synthesizing raw materials of DNA, mixing the phosphoramidite protected nucleotide monomer with an activator tetrazole to obtain a nucleoside phosphorous acid activated intermediate, wherein the 3' end of the nucleoside phosphorous acid activated intermediate is activated, and the 5' -hydroxyl is still protected by DMT and is subjected to condensation reaction with free 5' -hydroxyl in the solution.
And thirdly, carrying out a capping reaction, wherein a few 5' -hydroxyl groups possibly do not participate in the condensation reaction, stopping the reaction by using acetic anhydride and 1-methylimidazole, and then continuing the reaction, wherein the short segment can be separated in a subsequent link by selecting a purification mode according to the experimental requirements.
And fourthly, converting the phosphorylidene form into more stable phosphotriester under the action of oxidant iodine.
The utility model patent of patent No. CN202110205946.1 relates to nucleic acid synthesis system and includes two independent first kind pipeline and second kind pipeline, realizes the switching intercommunication of two kinds of pipeline states of circulation state and reinforced state through the circulation diverter valve. But the disadvantages are that: 1. a rotary valve is adopted, so that residues are always left and the cleaning is not easy to carry out; 2. a plurality of synthesis columns are used to achieve variable volumes, thereby increasing costs. 3. The first conduit is connected to the rotary valve for delivering multiple liquids simultaneously, and thus it is difficult to precisely control the flow rate.
SUMMERY OF THE UTILITY MODEL
To solve the above problems, the present invention provides a novel oligonucleotide synthesis system.
According to one aspect of the present invention, a novel oligo-nucleic acid synthesis system is provided, comprising a first pipeline, a second pipeline, a third pipeline, a circulation pipeline, a liquid inlet pipeline, a synthesis column, a liquid outlet pipeline and a bypass pipeline;
the first pipeline is connected with a plurality of phosphoramidite protected nucleotide monomer conveying pipelines and a detergent conveying pipeline through a 17-to-18-way valve;
the second pipeline is connected with a detergent conveying pipeline, a detritylation solvent conveying pipeline, an oxidant conveying pipeline and a cap A conveying pipeline through a 4-position 5-way valve;
the third pipeline is connected with a circulating cleaning waste discharge pipeline, a phosphorus deprotecting agent conveying pipeline, a circulating pipeline, a cap B conveying pipeline, a catalyst conveying pipeline, a vulcanizing agent conveying pipeline, an oxidant conveying pipeline, a detergent conveying pipeline and a detritylation solvent conveying pipeline through a 9-position 10-way valve;
the first pipeline, the second pipeline, the third pipeline, the liquid inlet pipeline and the bypass pipeline are connected through a 5-way valve;
the liquid inlet pipeline and the liquid outlet pipeline are connected with the synthetic column, and the circulating pipeline, the liquid outlet pipeline and the bypass pipeline are connected through a 2-position 3-way valve.
In some embodiments, the outlet of the 17-to-18 valve is connected to the inlet of the first conduit, and each inlet is connected to a plurality of the phosphoramidite protected nucleotide monomer delivery conduits and the detergent delivery conduit, respectively. Thus, the specific connection of the 17 to 18 way valve is described.
In some embodiments, the outlet end of the 4-to-5 way valve is connected to the inlet of the second conduit, and each inlet end is connected to the detergent delivery conduit, the detritylated solvent delivery conduit, the oxidant delivery conduit, and the cap a delivery conduit, respectively. Thus, a specific connection of the 4 to 5 way valve is described.
In some embodiments, the outlet end of the 9-position 10-way valve is connected to the inlet of the third pipeline, and each inlet end is connected to the circulation cleaning waste discharge pipeline, the phosphorus deprotecting agent conveying pipeline, the circulation pipeline, the cap B conveying pipeline, the catalyst conveying pipeline, the sulfidizing agent conveying pipeline, the oxidant conveying pipeline, the detergent conveying pipeline, and the detritylation solvent conveying pipeline, respectively. Thus, a specific connection of a 9 to 10 way valve is described.
In some embodiments, the ports of the 5-way valve are respectively connected with the outlet of the first pipeline, the outlet of the second pipeline, the outlet of the third pipeline, the inlet of the liquid inlet pipeline and the inlet of the bypass pipeline. Thus, a specific connection of the 5-way valve is described.
In some embodiments, the synthesis column has a synthesis chamber at the bottom and a hydraulic chamber at the top, and a piston column head is arranged between the synthesis chamber and the hydraulic chamber, and the hydraulic chamber is connected with a hydraulic system; the synthesis cavity is connected with an outlet of the liquid inlet pipeline, and the hydraulic cavity is connected with an inlet of the liquid outlet pipeline. Thus, the main structure of the synthesis column is described.
In some embodiments, the hydraulic system includes an oil tank, a first hydraulic oil delivery pipe, and a second hydraulic oil delivery pipe, where both the first hydraulic oil delivery pipe and the second hydraulic oil delivery pipe have one end communicating with the oil tank and the other end communicating with the hydraulic cavity. Thus, the structure of the hydraulic system is described.
In some embodiments, a hydraulic pump and a first pressure transmitter are disposed on the first hydraulic oil delivery pipe, and a hydraulic valve is disposed on the second hydraulic oil delivery pipe. Thus, the specific structure of the hydraulic system is further described.
In some embodiments, each end of the 2-to-3 way valve is connected to the outlet of the circulation pipeline, the outlet of the liquid outlet pipeline and the outlet of the bypass pipeline respectively. Thus, a specific connection of the 2 to 3 way valve is described.
In some embodiments, a peristaltic pump and a flow meter are disposed on each of the first, second, and third conduits. Thus, there is a peristaltic pump for powering each tube and a flow meter for detecting the flow rate for each tube.
Drawings
FIG. 1 is a schematic diagram of a novel oligonucleotide synthesis system according to an embodiment of the present invention;
FIG. 2 is a schematic diagram showing a partial structure of the oligonucleotide synthesis system shown in FIG. 1;
FIG. 3 is a schematic view of the structure of the synthesis column shown in FIG. 1.
In the figure: the device comprises a first pipeline 1, a second pipeline 2, a third pipeline 3, a circulating pipeline 4, a liquid inlet pipeline 5, a synthetic column 6, a liquid outlet pipeline 7, a bypass pipeline 8, a peristaltic pump 9, a phosphoramidite protected nucleotide monomer conveying pipeline 10, a 17-to-18-way valve 11, a detergent conveying pipeline 12,4-to-5-way valve 21, a detritylation solvent conveying pipeline 22, an oxidant conveying pipeline 23, a cap A conveying pipeline 24,9-to-10-way valve 31, a circulating cleaning waste discharge pipeline 32, a phosphorus deprotecting agent conveying pipeline 33, a cap B conveying pipeline 34, a catalyst conveying pipeline 35, a vulcanizing agent conveying pipeline 36,5-to-41,2-to-3-way valve 51, a second pressure transmitter 52, a synthetic cavity 61, a hydraulic cavity 62, an oil tank 63, a first hydraulic oil conveying pipeline 64, a second hydraulic oil conveying pipeline 65, a hydraulic pump 66, a hydraulic valve 67, a first pressure transmitter 68, a piston head 69, a UV detector 71, a UV detector 91 and a flowmeter.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings.
Fig. 1 schematically shows the structure of a novel oligo nucleic acid synthesis system according to an embodiment of the present invention, fig. 2 shows a part of the structure of the novel oligo nucleic acid synthesis system of fig. 1, and fig. 3 shows the structure of the novel synthesis column of fig. 1. As shown in fig. 1-3, the synthesis system mainly comprises a first pipeline 1, a second pipeline 2, a third pipeline 3, a circulation pipeline 4, a liquid inlet pipeline 5, a synthesis column 6, a liquid outlet pipeline 7, a bypass pipeline 8 and other structures.
The inlet of the first conduit 1 is connected to the outlet of a 17-to-18 way valve 11. Wherein, the inlet ends (17 in total) of the 17-to-18-way valves 11 are respectively connected with a plurality of phosphoramidite protected nucleotide monomer delivery pipelines 10 (generally 16) and detergent delivery pipelines 12 (generally 1).
The inlet of the second conduit 2 is connected to the outlet of a 4-to-5 way valve 21. The number of the inlet ends of the 4-to-5 way valve 21 is 4, which are respectively set as the a end, the b end, the c end and the d end, and the connection mode of each inlet end is as follows: the end a is connected with a detergent conveying pipeline 12, the end b is connected with a detritylation solvent conveying pipeline 22, the end c is connected with an oxidant conveying pipeline 23, and the end d is connected with a cap A conveying pipeline 24.
The inlet of the third conduit 3 is connected to the outlet of a 9-position 10-way valve 31. The 9-to-10 valve 31 has 9 inlet ports, which are respectively designated as an a port, a b port, a c port, a d port, an e port, an f port, a g port, an h port and an i port, and the inlet ports are connected as follows: the a end is connected with a circulation cleaning waste discharge pipeline 32, the B end is connected with a phosphorus deprotecting agent conveying pipeline 33, the c end is connected with the circulation pipeline 4, the d end is connected with a cap B conveying pipeline 34, the e end is connected with a catalyst conveying pipeline 35, the f end is connected with a vulcanizing agent conveying pipeline 36, the g end is connected with an oxidant conveying pipeline 23, the g end is connected with a detergent conveying pipeline 12, and the i end is connected with a detritylation solvent conveying pipeline 22.
The outlet of the first pipeline 1, the outlet of the second pipeline 2, the outlet of the third pipeline 3, the inlet of the liquid inlet pipeline 5 and the inlet of the bypass pipeline 8 are connected through ports of a 5-way valve 41. Wherein, the outlet of the first pipeline 1 is connected with 1 path of the 5-way valve 41, the outlet of the second pipeline 2 is connected with 2 paths of the 5-way valve 41, the outlet of the third pipeline 3 is connected with 3 paths of the 5-way valve 41, the inlet of the liquid inlet pipeline 5 is connected with 4 paths of the 5-way valve 41, and the inlet of the bypass pipeline 8 is connected with 5 paths of the 5-way valve 41.
Preferably, the peristaltic pump 9 and the flow meter 91 are arranged on the first pipeline 1, the second pipeline 2 and the third pipeline 3.
The outlet of the liquid inlet pipe 5 and the inlet of the liquid outlet pipe 7 are connected with the synthetic column 6, and the outlet of the circulating pipe 4, the outlet of the liquid outlet pipe 7 and the outlet of the bypass pipeline 8 are connected through the ports of a 2-to-3 valve 51.
Preferably, the second pressure transmitter 52 is arranged on the liquid inlet pipe 5, and the UV ultraviolet detector 71 is arranged on the liquid outlet pipe 7.
The bottom of the inner cavity of the synthetic column 6 is a synthetic cavity 61, the top is a hydraulic cavity 62, wherein the synthetic cavity 61 is connected with the outlet of the liquid inlet pipeline 5, and the hydraulic cavity 62 is connected with the inlet of the liquid outlet pipeline 7.
The hydraulic cavity 62 is connected with a hydraulic system, wherein the hydraulic system mainly comprises an oil tank 63, a first hydraulic oil conveying pipeline 64 and a second hydraulic oil conveying pipeline 65, the first hydraulic oil conveying pipeline 64 and the second hydraulic oil conveying pipeline 65 are both communicated with the oil tank 63 at one end, and the other end is communicated with the hydraulic cavity 62.
Preferably, a hydraulic pump 66 and a first pressure transmitter 67 are arranged on the first hydraulic oil supply line 64, and a hydraulic valve 68 is arranged on the second hydraulic oil supply line 65.
When the synthesis system is used to synthesize DNA, the operation and reaction process thereof include a plurality of steps, which are described in the following order.
A plunger post head 69 is provided between the synthesis chamber 61 and the hydraulic chamber 62, the plunger post head 69 dividing the inner chamber of the synthesis column 6 into the synthesis chamber 61 and the hydraulic chamber 62 and being capable of vertical movement along the inner chamber of the synthesis column 6 to adjust the volume of the synthesis chamber 61.
The specific method for adjusting the volume of the synthesis cavity 61 is as follows: when the solution is conveyed into the synthesis cavity 61, the hydraulic valve 68 on the second hydraulic oil conveying pipeline 65 is opened, the hydraulic oil in the hydraulic cavity 62 of the synthesis column 6 flows into the oil tank 63 through the second hydraulic oil conveying pipeline 65, the piston column head 69 moves upwards, and the volume of the synthesis cavity 61 is increased; when the solution in the synthesis chamber 61 is delivered, the hydraulic valve 68 on the second hydraulic oil delivery pipe 65 is closed, the hydraulic pump 66 on the first hydraulic oil delivery pipe 64 is opened, hydraulic oil flows into the hydraulic chamber 62 from the oil tank 63 through the first hydraulic oil delivery pipe 64, the piston column head 69 moves downwards, the volume of the synthesis chamber 61 is reduced, and the liquid in the synthesis chamber 61 can be discharged through the liquid outlet pipe 7.
When the synthesis system is used to synthesize DNA, the operation and reaction process thereof include a plurality of steps, which are described in the following order.
The first step is as follows: deblocking and detritylation.
The b-port of the 4-position 5-way valve 21 and the i-port of the 9-position 10-way valve 31 are opened, and the 2-way, 3-way and 4-way ports of the 5-way valve 41 are opened.
The detritylation solvent is introduced through the detritylation solvent delivery conduit 22, and then enters the second conduit 2 and the third conduit 3 through the 4-position 5-way valve 21 and the 9-position 10-way valve 31, respectively, and then enters the synthesis chamber 61 of the synthesis column 6 through the liquid inlet conduit 5. Wherein, the nucleotide with protected active group connected to the solid phase carrier reacts with dichloroacetic acid, the protecting group DMT of the 5' -hydroxyl group is removed, the free 5' -hydroxyl group is obtained, and finally the free 5' -hydroxyl group is discharged through a liquid outlet pipeline 7.
Preferably, the detritylation solvent may be dichloroacetic acid, trichloroacetic acid or toluene.
The second step: and (4) removing trityl and washing.
The b end of the 4 th 5-way valve 21 and the i end of the 9 th 10-way valve 31 are closed, and the a end of the 4 th 5-way valve 21 and the h end of the 9 th 10-way valve 31 are opened.
The detergent is introduced through the detergent conveying pipeline 12, then the detergent respectively enters the second pipeline 2 and the third pipeline 3 through the a end of the 4-position 5-way valve 21 and the h end of the 9-position 10-way valve 31, then enters the synthesis cavity 61 of the synthesis column 6 through the liquid inlet pipeline 5, and finally is discharged through the liquid outlet pipeline 7.
Preferably, the detergent may employ acetonitrile.
The third step: coupling and activating.
The a end of the 4-position 5-way valve 21 and the h end of the 9-position 10-way valve 31 are closed, the inlet ends of the 17-position 18-way valve 11 are opened, and the e end of the 9-position 10-way valve 31 and the 1, 3 and 4 ways of the 5-way valve 41 are opened.
A plurality of phosphoramidite protected nucleotide monomers are introduced through the 10 conveying pipelines of each phosphoramidite protected nucleotide monomer, then enter the first pipeline 1 through the 17 th 18 th through valve 11, and simultaneously, the catalyst is introduced through the 35 conveying pipelines of the catalyst, then enter the third pipeline 3 through the 9 th 10 th through valve 31, so that the plurality of phosphoramidite protected nucleotide monomers and the catalyst can enter the synthesis cavity 61 of the synthesis column 6 through the liquid inlet pipeline 5, so that the catalytic reaction is carried out, and a nucleoside phosphorous acid activated intermediate is obtained, wherein the 3' end of the nucleoside phosphorous acid activated intermediate is activated, the 5' -hydroxyl is still protected by DMT, and the nucleoside phosphorous acid activated intermediate and the free 5' -hydroxyl in the synthesis cavity 61 of the synthesis column 6 are subjected to condensation reaction.
The fourth step: and (4) coupling circulation.
Closing the inlet ends of the 17-position 18-way valve 11, the e end of the 9-position 10-way valve 31 and the 1 way, the 3 way and the 4 way of the 5-way valve 41; then the c-port of the 9-position 10-way valve 31 and the 3-way and 4-way of the 5-way valve 41 are opened.
At this time, the circulation pipeline 4 is communicated with the third pipeline 3, and the nucleoside phosphite activated intermediate can circularly flow through the circulation pipeline 4, the third pipeline 3, the liquid inlet pipeline 5 and the synthesis cavity 61 of the synthesis column 6, so that the full reaction can be carried out.
The fifth step: and (6) washing.
The c-port of the 9-position 10-way valve 31 and the 3-and 4-ports of the 5-way valve 41 are closed, and then the 17-position 18-way valve 11 is opened again with the inlet end of the detergent delivery pipe 12, the 1-port, 3-and 4-ports of the 5-way valve 41 and the h-port of the 9-position 10-way valve 31.
Acetonitrile serving as a detergent is respectively introduced into the first pipeline 1 and the third pipeline 3 through a detergent conveying pipeline 12, then enters the synthesis cavity 61 of the synthesis column 6 through a liquid inlet pipeline 5 for washing, and finally is discharged through a liquid outlet pipeline 7.
And a sixth step: and (5) cleaning a circulating pipeline.
The a end of the 9 position 10 way valve 31 and the valve between the outlet pipe 7 and the circulation pipe 4 on the 2 position 3 way valve 51 are opened.
At this time, the detergent may enter the exhaust line 32 through the circulation line 4 and wash the circulation line 4.
The seventh step: and (4) vulcanizing.
The inlet end of the 17 th 18-way valve 11 from the detergent delivery pipe 12, the 1 st, 3 rd and 4 th ways of the 5 th way valve 41 and the a and h ends of the 9 th 10-way valve 31 are closed, and then the f end of the 9 th 10-way valve 31 and the 3 rd and 4 th ways of the 5 th way valve 41 are opened.
And (3) introducing the vulcanizing reagent through the vulcanizing reagent conveying pipeline 36, and enabling the vulcanizing reagent to sequentially pass through the third pipeline 3, the liquid inlet pipeline 5, the hydraulic cavity 62 of the synthetic column 6 and the liquid outlet pipeline 7 so as to perform a vulcanizing reaction.
Eighth step: and (4) sulfurizing and washing.
The f-port of the 9-to-10 way valve 31 is closed and the h-port of the 9-to-10 way valve 31 is opened.
Acetonitrile as a detergent is introduced through the detergent delivery pipe 12, and then passes through the third pipe 3, the liquid inlet pipe 5, the hydraulic chamber 62 of the synthesis column 6, and the liquid outlet pipe 7, so that washing is performed.
The ninth step: and (4) end capping.
The h end of the 9-position 10-way valve 31 and the 3-way and 4-way of the 5-way valve 41 are closed, and the d end of the 4-position 5-way valve 21, the d end of the 9-position 10-way valve 31 and the 2-way, 3-way and 4-way of the 5-way valve 41 are opened.
The cap A and the cap B are respectively introduced into the second pipeline 2 and the third pipeline 3 through the cap A conveying pipeline 24 and the cap B conveying pipeline 34, and then the cap A and the cap B can continuously enter the hydraulic cavity 62 of the synthetic column 6 through the liquid inlet pipeline 5 and finally enter the liquid outlet pipeline 7, so that the end capping effect is realized.
The tenth step: and (4) end-capping circulation.
The d end of the 4-position 5-way valve 21, the d end of the 9-position 10-way valve 31 and the 2-way of the 5-way valve 41 are closed, and then the c end of the 9-position 10-way valve 31 and the valve between the liquid outlet pipeline 7 and the circulating pipeline 4 on the 2-position 3-way valve 51 are opened.
In addition, the cap a and the cap B circulate among the third pipe 3, the liquid inlet pipe 5, the hydraulic chamber 62 of the synthesis column 6, the liquid outlet pipe 7, and the circulation pipe 4.
The eleventh step: and (5) end-capping and washing.
The c end of the 9-position 10-way valve 31 and the valve between the liquid outlet pipeline 7 and the circulating pipeline 4 on the 2-position 3-way valve 51 are closed, and the a end of the 4-position 5-way valve 21, the 1 way, the 3 way and the 4 way of the 5-way valve 41 and the h end of the 9-position 10-way valve 31 are opened.
Acetonitrile as a detergent is introduced through the detergent conveying pipeline 12, and then the acetonitrile respectively enters the second pipeline 2 and the third pipeline 3, then enters the synthesis cavity 61 of the synthesis column 6 through the liquid inlet pipeline 5, and finally is discharged through the liquid outlet pipeline 7, so that the end-capping washing effect is realized.
The twelfth step: and (5) cleaning the circulating pipe.
The valve between the liquid outlet pipe 7 and the circulating pipe 4 on the 2 position 3 way valve 51 and the branch a end of the 9 position 10 way valve 31 are opened.
At this time, the introduced detergent is introduced into the circulation cleaning exhaust line 32 through the circulation line 4 to be cleaned.
And a thirteenth step of: and (4) removing phosphorus.
The a end of the 4-to-5 way valve 21, the 1 way, the 3 way and the 4 way of the 5 way valve 41, the a end and the h end of the 9-to-10 way valve 31 and the valve between the liquid outlet pipeline 7 and the circulating pipeline 4 on the 2-to-3 way valve 51 are closed, and then the b end of the 9-to-10 way valve 31 and the 3 way and the 4 way of the 5 way valve 41 are opened.
The phosphorus deprotecting agent is introduced into the third pipeline 3 through the phosphorus deprotecting agent conveying pipeline 33, enters the synthesis cavity 61 of the synthesis column 6 along the liquid inlet pipeline 5, and is finally discharged through the liquid outlet pipeline 7, so as to realize the phosphorus deprotecting effect.
A fourteenth step of: and (4) carrying out phosphorus deprotection cycle.
The b end of the 9-position 10-way valve 31 is closed, and the c end of the 9-position 10-way valve 31 and a valve between the liquid outlet pipeline 7 and the circulating pipeline 4 on the 2-position 3-way valve 51 are opened.
At this time, the introduced phosphorus deprotecting agent circulates among the third pipe 3, the liquid inlet pipe 5, the hydraulic chamber 62 of the synthesis column 6, the liquid outlet pipe 7 and the circulation pipe 4.
The fifteenth step: and (5) finally washing.
The c end of the 9-position 10-way valve 31 and the valve between the liquid outlet pipeline 7 and the circulating pipeline 4 on the 2-position 3-way valve 51 are closed, and then the 17-position 18-way valve 11 is opened to connect the inlet end with the detergent conveying pipeline 12, the 1 path, the 2 path, the 3 path and the 4 path of the 5-way valve 41, the a end of the 4-position 5-way valve 21 and the h end of the 9-position 10-way valve 31.
Acetonitrile serving as a detergent is introduced through a detergent conveying pipeline 12, then the acetonitrile can enter a first pipeline 1, a second pipeline 2 and a third pipeline 3 respectively, then enters a synthesis cavity 61 of a synthesis column 6 through a liquid inlet pipeline 5, and is finally discharged through a liquid outlet pipeline 7, so that the effect of finally washing all pipelines of the system is achieved.
Sixteenth, step: and (5) cleaning a circulating pipeline.
The valve between the outlet pipe 7 and the circulation pipe 4 of the 9 th 10 way valve 31 and the 2 nd 3 way valve 51 is opened.
And the detergent is continuously introduced from the detergent conveying pipeline 12, enters the synthesis cavity 61 of the synthesis column 6 through the liquid inlet pipeline 5 after entering the first pipeline 1, the second pipeline 2 and the third pipeline 3 respectively, then passes through the liquid outlet pipeline 7 and the circulating pipeline 4, and finally is discharged through the circulating cleaning waste discharge pipeline 32.
After the above steps are completed, the above steps may be repeated in accordance with the base sequence of the nucleic acid until the whole DNA synthesis work is completed.
When the solution does not need to pass through the synthesis column 6, the 5 end of the 4-to-5 valve 21 can be opened, and the solution directly enters the liquid outlet pipeline 7 through the bypass pipeline 8 and is discharged.
What has been described above are only some embodiments of the invention. For those skilled in the art, without departing from the inventive concept, several modifications and improvements can be made, which are within the scope of the invention.

Claims (10)

1. A novel oligonucleotide synthesis system, characterized in that: comprises a first pipeline (1), a second pipeline (2), a third pipeline (3), a circulating pipeline (4), a liquid inlet pipeline (5), a synthesizing column (6), a liquid outlet pipeline (7) and a bypass pipeline (8);
the first pipeline (1) is connected with a plurality of phosphoramidite protected nucleotide monomer conveying pipelines (10) and a detergent conveying pipeline (12) through a 17-to-18-way valve (11);
the second pipeline (2) is connected with a detergent conveying pipeline (12), a detritylation solvent conveying pipeline (22), an oxidant conveying pipeline (23) and a cap A conveying pipeline (24) through a 4-position 5-way valve (21);
the third pipeline (3) is connected with a circulating cleaning waste discharge pipeline (32), a phosphorus deprotecting agent conveying pipeline (33), a circulating pipeline (4), a cap B conveying pipeline (34), a catalyst conveying pipeline (35), a vulcanizing agent conveying pipeline (36), an oxidant conveying pipeline (23), a detergent conveying pipeline (12) and a trityl-removed solvent conveying pipeline (22) through a 9-to-10-way valve (31);
the first pipeline (1), the second pipeline (2), the third pipeline (3), the liquid inlet pipeline (5) and the bypass pipeline (8) are connected through a 5-way valve (41);
liquid inlet pipe (5) with liquid outlet pipe (7) all with synthetic column (6) are connected, circulating line (4) liquid outlet pipe (7) with side branch's pipeline (8) are connected through 2 to 3 logical valves (51).
2. The novel oligonucleotide synthesis system of claim 1, wherein: the outlet end of the 17-to-18 way valve (11) is connected with the inlet of the first pipeline (1), and each inlet end is respectively connected with a plurality of phosphoramidite protected nucleotide monomer conveying pipelines (10) and the detergent conveying pipeline (12).
3. The novel oligonucleotide synthesis system of claim 1, wherein: the outlet end of the 4-position 5-way valve (21) is connected with the inlet of the second pipeline (2), and each inlet end is respectively connected with the detergent conveying pipeline (12), the detritylation solvent conveying pipeline (22), the oxidant conveying pipeline (23) and the cap A conveying pipeline (24).
4. The novel oligonucleotide synthesis system of claim 1, wherein: the outlet end of the 9-position 10-way valve (31) is connected with the inlet of the third pipeline (3), and each inlet end is respectively connected with the circulating cleaning waste discharge pipeline (32), the phosphorus deprotecting agent conveying pipeline (33), the circulating pipeline (4), the cap B conveying pipeline (34), the catalyst conveying pipeline (35), the sulfuration reagent conveying pipeline (36), the oxidant conveying pipeline (23), the detergent conveying pipeline (12) and the detritylation solvent conveying pipeline (22).
5. The novel oligonucleotide synthesis system of claim 1, wherein: and each path of port of the 5-way valve (41) is respectively connected with the outlet of the first pipeline (1), the outlet of the second pipeline (2), the outlet of the third pipeline (3), the inlet of the liquid inlet pipeline (5) and the inlet of the bypass pipeline (8).
6. The novel oligonucleotide synthesis system of claim 1, wherein: the synthetic column (6) is provided with a synthetic cavity (61) at the bottom and a hydraulic cavity (62) at the top, a piston column head (69) is arranged between the synthetic cavity (61) and the hydraulic cavity (62), and the hydraulic cavity (62) is connected with a hydraulic system; the synthesis cavity (61) is connected with an outlet of the liquid inlet pipeline (5), and the hydraulic cavity (62) is connected with an inlet of the liquid outlet pipeline (7).
7. The novel oligonucleotide synthesis system of claim 6, wherein: the hydraulic system comprises an oil tank (63), a first hydraulic oil conveying pipeline (64) and a second hydraulic oil conveying pipeline (65), wherein the first hydraulic oil conveying pipeline (64) and the second hydraulic oil conveying pipeline (65) are both one ends and communicated with the oil tank (63), and the other ends are communicated with the hydraulic cavity (62).
8. The novel oligonucleotide synthesis system of claim 7, wherein: the first hydraulic oil conveying pipeline (64) is provided with a hydraulic pump (66) and a first pressure transmitter (67), and the second hydraulic oil conveying pipeline (65) is provided with a hydraulic valve (68).
9. The novel oligonucleotide synthesis system of claim 1, wherein: and each end of the 2-position 3-way valve (51) is respectively connected with the circulating pipeline (4), the outlet of the liquid outlet pipeline (7) and the outlet of the bypass pipeline (8).
10. A novel oligonucleotide synthesis system according to any one of claims 1-9, wherein: the first pipeline (1), the second pipeline (2) and the third pipeline (3) are all provided with a peristaltic pump (9) and a flow meter (91).
CN202221346930.9U 2022-05-31 2022-05-31 Novel oligonucleotide synthesis system Active CN218077815U (en)

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