CN117384749A - Method for loading sequencing liquid onto sequencing slide, flow path system for gene sequencing and gene sequencer - Google Patents
Method for loading sequencing liquid onto sequencing slide, flow path system for gene sequencing and gene sequencer Download PDFInfo
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- 238000012163 sequencing technique Methods 0.000 title claims abstract description 168
- 239000007788 liquid Substances 0.000 title claims abstract description 31
- 238000000034 method Methods 0.000 title claims abstract description 14
- 108090000623 proteins and genes Proteins 0.000 title claims description 40
- 239000000523 sample Substances 0.000 claims abstract description 120
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 85
- 239000012472 biological sample Substances 0.000 claims abstract description 48
- 230000002068 genetic effect Effects 0.000 claims abstract description 9
- 239000003795 chemical substances by application Substances 0.000 claims description 10
- 239000012530 fluid Substances 0.000 claims description 10
- 238000010926 purge Methods 0.000 claims description 10
- 238000005406 washing Methods 0.000 claims description 5
- 239000012459 cleaning agent Substances 0.000 abstract description 29
- 238000005086 pumping Methods 0.000 description 49
- 238000004891 communication Methods 0.000 description 18
- 238000004140 cleaning Methods 0.000 description 17
- 239000002699 waste material Substances 0.000 description 15
- 238000010586 diagram Methods 0.000 description 12
- 230000000694 effects Effects 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 2
- 238000006467 substitution reaction Methods 0.000 description 2
- GNFTZDOKVXKIBK-UHFFFAOYSA-N 3-(2-methoxyethoxy)benzohydrazide Chemical compound COCCOC1=CC=CC(C(=O)NN)=C1 GNFTZDOKVXKIBK-UHFFFAOYSA-N 0.000 description 1
- YTAHJIFKAKIKAV-XNMGPUDCSA-N [(1R)-3-morpholin-4-yl-1-phenylpropyl] N-[(3S)-2-oxo-5-phenyl-1,3-dihydro-1,4-benzodiazepin-3-yl]carbamate Chemical compound O=C1[C@H](N=C(C2=C(N1)C=CC=C2)C1=CC=CC=C1)NC(O[C@H](CCN1CCOCC1)C1=CC=CC=C1)=O YTAHJIFKAKIKAV-XNMGPUDCSA-N 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 238000011109 contamination Methods 0.000 description 1
- 238000007872 degassing Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 230000002572 peristaltic effect Effects 0.000 description 1
- 238000000638 solvent extraction Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
- C12M33/06—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles for multiple inoculation or multiple collection of samples
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6869—Methods for sequencing
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Abstract
The present invention relates to a method of loading a sequencing liquid to a genetic sequencing slide, the genetic sequencing slide comprising a plurality of sample channels, the method comprising independently loading a plurality of sample channels with a sequencing liquid, the sequencing liquid comprising a biological sample, a sequencing reagent or a cleaner; independent loading of biological samples, sequencing reagents or cleaning agents in at least two sample channels is thereby achieved.
Description
Technical Field
The invention relates to the technical field of gene sequencers, in particular to a method for loading sequencing liquid to a sequencing slide, a flow path system for gene sequencing and a gene sequencer.
Background
The test slide in the gene sequencer is divided into at least two sample channels, each of which can theoretically be loaded with a different biological sample. However, due to the complexity of the fluid system, the existing device does not have the function of loading different biological samples in each sample channel, which is completed by a special loading tool, and can not automatically use a plurality of different cleaning agents to clean each flow path component, so that the cleaning work efficiency is low, and time and labor are wasted.
Disclosure of Invention
The invention provides a method for loading a sequencing liquid to a gene sequencing slide, wherein the gene sequencing slide comprises a plurality of sample channels, the method comprises independently loading the sequencing liquid to the plurality of sample channels, and the sequencing liquid comprises a biological sample, a sequencing reagent or a cleaning agent.
In some embodiments, the step of loading the sequencing liquid comprises allowing the sequencing liquid to selectively flow in from one of the two ends of the sample channel and out from the other of the two ends of the sample channel, such that the sequencing liquid can flow in from one of the two ends of the sample channel and also flow in from the other of the two ends of the sample channel.
In some embodiments, the step of loading the sequencing liquid comprises flowing the sequencing liquid through the sample channel by positive or negative pressure.
The present invention also provides a flow path system for gene sequencing, comprising:
a genetic sequencing slide having a plurality of sample channels;
at least one of the following:
a biological sample loading flow path configured to be able to independently load a biological sample to the plurality of sample channels;
a sequencing reagent loading flow path configured to be capable of independently loading sequencing reagents into a plurality of sample channels; and
and a washing channel configured to be able to independently load washing agents to the plurality of sample channels.
In some embodiments, the flow path system comprises at least one of:
a biological sample loading flow path configured to enable a biological sample to flow into the sample channel from one of both ends of the sample channel and to flow out from the other of both ends of the sample channel, so that the biological sample can flow into the sample channel from one end of the sample channel and also can flow into the sample channel from the other end of the sample channel;
a sequencing reagent loading flow path configured to enable a sequencing reagent to flow selectively into the sample channel from one of the two ends of the sample channel and out of the other of the two ends of the sample channel, such that the sequencing reagent can flow into the sample channel from one end of the sample channel and also can flow into the sample channel from the other end of the sample channel; and
the cleaning flow path is configured to enable the cleaning agent to flow into the sample channel from one of both ends of the sample channel selectively, and flow out from the other of both ends of the sample channel, so that the cleaning agent can flow into the sample channel from one end of the sample channel and also can flow into the sample channel from the other end of the sample channel.
In some embodiments, the flow path system comprises at least one of:
a biological sample loading flow path configured to flow a biological sample through a sample channel by positive or negative pressure;
a sequencing reagent loading flow path configured to flow a sequencing reagent through the sample channel by positive or negative pressure; and
a purge flow path configured to flow a purge through the sample channel by positive or negative pressure.
In some embodiments, the flow path system includes a plurality of gene sequencing slides that operate independently of one another.
In some embodiments, one end of each of the plurality of sample channels in one genetic sequencing slide is fluidically pooled to one port, such that sequencing fluid can flow into or out of the sample channels through the port.
In some embodiments, the invention also provides a gene sequencer, using the method of loading a sequencing fluid onto a gene sequencing slide.
In some embodiments, the invention also provides a gene sequencer comprising the flow path system for gene sequencing.
According to the method and the flow path system for loading the sequencing liquid to the sequencing slide, at least one of the following technical effects can be achieved: independently loading biological samples in a plurality of sample channels of a sequencing slide, so that sequencing requirements of different biological samples are better met; separately loading sequencing reagents into a plurality of sample channels, respectively, to meet the need for using different sequencing reagents in sequencing; and cleaning the plurality of sample channels in the flow path system independently by using cleaning agents respectively so as to meet different cleaning requirements of each sample channel.
Drawings
FIG. 1 is a first exemplary diagram of a flow path system for gene sequencing of the present invention;
FIG. 2 is a second exemplary diagram of a fluidic system for gene sequencing according to the present invention;
FIG. 3 is a third exemplary diagram of a fluidic system for gene sequencing according to the present invention;
FIG. 4 is a fourth exemplary diagram of a fluidic system for gene sequencing according to the present invention;
FIG. 5 is a fifth exemplary diagram of a fluidic system for gene sequencing according to the present invention;
FIG. 6 is a sixth exemplary diagram of a fluidic system for gene sequencing according to the present invention;
FIG. 7 is a seventh exemplary illustration of a fluidic system for gene sequencing according to the present invention;
FIG. 8 is an eighth exemplary diagram of a fluidic system for gene sequencing according to the present invention;
FIG. 9 is a ninth exemplary diagram of a fluidic system for gene sequencing according to the present invention;
FIG. 10 is a tenth exemplary diagram of a fluidic system for gene sequencing according to the present invention;
FIG. 11 is an eleventh illustration of a fluidic system for gene sequencing according to the present invention;
FIG. 12 is a diagram of a twelfth example of a fluidic system for gene sequencing according to the present invention;
FIG. 13 is a thirteenth exemplary illustration of a fluidic system for gene sequencing according to the present invention;
FIG. 14 is a fourteenth exemplary illustration of a fluidic system for gene sequencing according to the present invention;
FIG. 15 is a fifteenth exemplary illustration of a fluidic system for gene sequencing according to the present invention; and
FIG. 16 is a sixteenth exemplary diagram of a fluidic system for gene sequencing according to the present invention.
Detailed Description
The following detailed description of the invention is made in connection with specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention.
Loading flow path
The flow path system 10 for gene sequencing of the present invention comprises a biological sample region 11, a first reversing valve assembly 12, a sequencing slide 13 and a suction device 14, wherein the biological sample region 11 is provided with a plurality of sample storage bins respectively containing different biological samples, the sequencing slide 13 is provided with a plurality of mutually independent sample channels, the first reversing valve assembly 12 is respectively communicated with the biological sample region 11, the sequencing slide 13 and the suction device 14, and the suction device 14 can be communicated between the biological sample region 11 and the sequencing slide 13 in a switchable manner through the first reversing valve assembly 12 so as to realize the suction and the transmission of different biological samples into the plurality of sample channels of the sequencing slide 13 respectively. The biological sample area 11 can be divided into several sample storage compartments independent of each other as required, the separation contour lines not being shown in the drawing.
The invention will be described below with four components as examples, it being understood that the above embodiments of the invention may also be selected from more or fewer components, including at least the sample storage compartment, the reversing valve, the pumping components of the pumping device, the sample channels, etc., as well as from more or fewer components, and optionally loading biological samples onto some of the sample channels, where "plurality" means at least two, and "first" and "second" etc. are used merely for literal distinction, without any limitation, "communicating" means communicating in a flow path.
In some embodiments, as shown in fig. 1, a first to fourth sample storage bins for respectively accommodating first to fourth biological samples are provided in the biological sample region 11, the first reversing valve assembly 12 includes four first reversing valves T1-T4, the sequencing slide 13 is provided with four first to fourth sample channels C1-C4 independent of each other, the aspiration device 14 includes first to fourth aspiration components P1-P4, the first ports of the four first reversing valves T1-T4 are respectively communicated with the first to fourth sample storage bins through four sample needles through four first lines L1, the second ports of the four first reversing valves T1-T4 are respectively communicated with the I ports (Inlet) of the first to fourth aspiration components P1-P4 through four second lines L2, the third ports of the four first reversing valves T1-T4 are respectively communicated with the respective Inlet ends of the first to fourth sample channels C1-C4 through four third lines L3, and a waste liquid flow control valve is provided between the first to fourth sample channels C1-C4 and the outlet ends of the sample region 16. Alternatively, the first reversing valve assembly 12 includes four three-way solenoid valves and a base manifold. Alternatively, the suction device 14 comprises four suction pumps, each suction pump comprising a syringe and a valve head, the syringe being in selective communication with one of the multiple ports of the valve head.
In some embodiments, different biological samples are placed in the first through fourth sample storage bins, respectively.
In some embodiments, as shown in fig. 1, the four first reversing valves T1-T4 are powered, i.e., the first ports of the four first reversing valves T1-T4 are respectively in communication with the second port, the second port is disconnected from the third port, the I ports of the first through fourth pumping sections P1-P4 provide negative pressure to pump the first through fourth samples from the first through fourth sample storage bins, respectively, through the first line L1, the four first reversing valves T1-T4, the second line L2 to the I ports of the first through fourth pumping sections P1-P4, and finally the biological samples (e.g., DNB1-DNB 4) are temporarily stored in the four second lines L2, respectively.
In some embodiments, as shown in FIG. 2, the four first reversing valves T1-T4 are de-energized, the second ports of the four first reversing valves T1-T4 are respectively in communication with the third port, the first port and the second port are disconnected, the first through fourth pumping sections P1-P4 provide positive pressure to push each biological sample temporarily stored in the second line L2, the redundant biological sample is discharged from the outlet ends of the first through fourth sample channels C1-C4 through the four first reversing valves T1-T4, the third line L3, the first through fourth sample channels C1-C4, and the waste fluid zone 16 through the rotary valve 15.
In some embodiments, as shown in fig. 3, the flow path system for gene sequencing of the present invention includes a sequencing reagent zone 17 storing a sequencing reagent, the first reversing valve assembly 12 includes a second reversing valve T5, a first port of the second reversing valve T5 communicates with a first sequencing reagent needle 171 of the sequencing reagent zone 17, the first sequencing reagent needle 171 communicates with a sequencing reagent such as an auxiliary reagent, a second port of the second reversing valve T5 communicates with ports B of the first to fourth pumping sections P1 to P4, and a third port of the second reversing valve T5 communicates with outlet ends of the first to fourth sample channels C1 to C4 and the rotary valve 15, respectively, through a tee. The sequencing reagent zone 17 may be partitioned into separate reagent reservoirs as desired, the partitioning contours not being shown in the figures.
In some embodiments, as shown in FIG. 3, the four first reversing valves T1-T4 are de-energized, the second reversing valve T5 is energized, and the first through fourth pumping sections P1-P4 provide negative pressure such that sequencing reagents enter the first through fourth pumping sections P1-P4 from sequencing reagent zone 17 through second reversing valve T5 and port B of first through fourth pumping sections P1-P4 for temporary storage to vent air in the tubing; the O-ports (Outlet) of the first to fourth pumping sections P1 to P4 communicate with the waste liquid region 16 through four fourth lines L4.
In some embodiments, as shown in FIG. 4, when the first through fourth pumping sections P1-P4 are switched to the O-port, the first through fourth pumping sections P1-P4 provide a positive pressure pushing force to discharge sequencing reagents in the temporary storage first through fourth pumping sections P1-P4 into the waste region 16 through the fourth line L4.
In some embodiments, four first reversing valves T1-T4 are energized, the second reversing valve T5 is de-energized, the first through fourth pumping sections P1-P4 are switched to port I (Inlet), positive pressure is provided to push sequencing reagents temporarily in the first through fourth pumping sections P1-P4 through the second line L2, four first reversing valves T1-T4, the first line L1 to each sample needle of the biological sample region 11, such that the first and second lines L1 and L2 are filled with sequencing reagents, air in the lines is expelled, and the flow path is referenced to FIG. 1.
In some embodiments, the four first reversing valves T1-T4 are de-energized, the second reversing valve T5 is energized, the rotary valve 15 is switched to a hole site in communication with the waste region 16, the first through fourth pumping sections P1-P4 are switched to an I port, positive pressure is provided to push the sequencing reagent held temporarily in the first through fourth pumping sections P1-P4, sequentially through the second line L2, the four first reversing valves T1-T4, the third line L3, the first through fourth sample channels C1-C4, the fifth line L5, the rotary valve 15, and finally into the waste region 16. This replaces the old sequencing reagent in the first through fourth sample channels C1-C4, the replaced old sequencing reagent and the excess sequencing reagent drain from the outlet ends of the first through fourth sample channels C1-C4 to the waste region 16, preventing contamination of the sequencing reagent in the first through fourth sample channels C1-C4 during subsequent testing steps, and the flow diagram is similar to FIG. 2.
In some embodiments, the biological sample is added only to the fourth biological storage bin.
In some embodiments, as shown in fig. 5, a fifth reversing valve T9 is disposed on the first line L1 between the first reversing valve T4 and the fourth sample storage compartment, a first port of the fifth reversing valve T9 is in communication with the rotary valve 15, a second port of the fifth reversing valve T9 is in communication with the fourth sample storage compartment, and a third port of the fifth reversing valve T9 is in communication with the first port of the first reversing valve T4. In the case where the four first reversing valves T1 to T4 and T9 are powered on and the second reversing valve T5 is powered off, the fourth suction means P4 provides negative pressure to suck the fourth biological sample from the fourth sample storage bin, sequentially through the sample needle, the fifth reversing valve T9, the rotary valve 15 and the second reversing valve T5 to the ports B of the first to fourth suction means P1 to P4, and at this time the fifth line L5 from the rotary valve 15 to the outlet ends of the first to fourth sample channels C1 to C4 is filled with the fourth biological sample, thereby washing and filling the line with the biological sample.
In some embodiments, as shown in fig. 6, the fifth reversing valve T9 is powered, the four first reversing valves T1-T4 are powered off, the second reversing valve T5 is powered on, the first through fourth pumping components P1-P4 are switched to port I, the fourth biological sample in the fifth line L5 is provided to be pumped under negative pressure from the outlet end of the sequencing slide 13 into the first through fourth biological sample channels C1-C4, respectively, the excess fourth biological sample is discharged from the inlet end of the sequencing slide 13, through the third line L3, the four first reversing valves T1-T4, the second line L2, through the port I of the first through fourth pumping components P1-P4 into the first through fourth pumping components P1-P4, thereby completing the loading of the fourth biological sample into the first through fourth sample channels C1-C4. Referring to fig. 4, the first to fourth pumping sections P1 to P4 are switched to the O-port, and positive pressure is provided to push the biological sample in the first to fourth pumping sections P1 to P4 to be discharged into the waste liquid region 16 through the fourth line L4.
In some embodiments, as shown in fig. 7, the sequencing reagent zone 17 is further provided with a plurality of second sequencing reagent pins 172, each corresponding to a plurality of different sequencing reagents. The rotary valve 15 is provided with a plurality of hole sites, which are respectively communicated with a plurality of second sequencing reagent pins 172 one by one, and when the rotary valve 15 is switched to one of the hole sites, the rotary valve 15 is communicated with one of the second sequencing reagent pins 172 corresponding to the hole site.
In some embodiments, as shown in FIG. 7, a sequencing reagent pretreatment device 24 is provided on the line between the plurality of second sequencing reagent pins 172 and the rotary valve 15, having at least one of filtering, removing dissolved oxygen, degassing, etc., the sequencing reagents withdrawn from the sequencing reagent zone 17.
There are two ways to deliver sequencing reagents: bypass and bychp.
The Bypass operation steps are as follows:
(1) As shown in fig. 7, the four first reversing valves T1-T4 are energized, the second reversing valve T5 is de-energized, the rotary valve 15 is switched to communicate with the desired sequencing reagent through one second sequencing reagent needle 172, the first through fourth pumping sections P1-P4 are switched to port B, the desired sequencing reagent is pumped from the sequencing reagent zone 17 by providing negative pressure, and the desired sequencing reagent is finally introduced into the first through fourth pumping sections P1-P4 through the second sequencing reagent needle 172, the rotary valve 15, the fifth line L5, the second reversing valve T5, port B of the first through fourth pumping sections P1-P4;
(2) The sequencing reagent can also enter the first to fourth pumping parts P1 to P4 or exceed the second reversing valve T5, so long as the condition that the sequencing reagent completely fills the fifth pipeline L5 is satisfied;
(3) The first through fourth pumping sections P1-P4 switch to the O-port, providing positive pressure to transfer sequencing reagents from the first through fourth pumping sections P1-P4 to the waste region 16 via the fourth line L4.
The operation steps of Bychip are as follows:
(1) As shown in fig. 8, the four first reversing valves T1-T4 are de-energized, the second reversing valve T5 is energized, the first rotary valve 15 is switched to communicate with the desired sequencing reagent through one second sequencing reagent needle 172, the first through fourth pumping sections P1-P4 are switched to the I port, the first through fourth pumping sections P1-P4 provide negative pressure to introduce the desired sequencing reagent into the biological channels C1-C4 through the second sequencing reagent needle 172 rotary valve 15, the fifth channel L5 and the outlet end of the sequencing slide 13, the old sequencing reagent and the redundant new sequencing reagent are removed from the inlet end of the sequencing slide 13, through the third pipeline L3, the first through fourth pumping sections P1-P4, the second pipeline L2 and the I port of the first through fourth pumping sections P1-P4, and into the first through fourth pumping sections P1-P4.
(2) The first to fourth suction parts P1 to P4 switch O-ports, and provide positive pressure pushing force to push out the sequencing reagent therein and discharge the sequencing reagent into the waste liquid area 16 through the fourth pipeline L4.
One of the following operational steps may be taken:
1. executing all steps of the Bypass, and executing all steps of the Bychip;
2. executing the steps (1) and (2) of Bypass, and executing the steps (1) and (2) of Bychip;
steps (1) and (2) of Bypass are performed, followed by step (1) of Bypass, and rotary valve 15 is switched to communicate with another desired sequencing reagent, followed by steps (1) and (2) of Bypass.
According to the loading flow path of the present invention, at least one of the following technical effects can be brought about:
different biological samples are respectively and independently loaded in a plurality of sample channels of the same sequencing slide, so that the sequencing requirements of the different biological samples are better met, and the flexibility of gene sequencing is improved;
a plurality of different sequencing reagents are selectively loaded into selected sample channels, respectively, to meet the need for using different sequencing reagents in different channels in gene sequencing.
Cleaning flow path
In some embodiments, as shown in fig. 9, the flow path system for gene sequencing further comprises a wash flow path comprising a wash zone 18 and a second valve block 19, the wash zone 18 storing a plurality of different types of wash agents, e.g. comprising a first wash agent W1, a second wash agent W2, a third wash agent W3, a fourth wash agent W4, a fifth wash agent W5, the second valve block 18 comprising a plurality of inlets and a common outlet, the plurality of inlets respectively communicating with the plurality of different types of wash agents, e.g. the second valve block 19 comprising four two-way valves T10-T13, each two-way valve having an inlet, each inlet communicating with a respective wash agent, the outlets of each two-way valve being connected to form a common outlet and being in selective communication with the second reversing valve T5 and the outer walls of the plurality of second sequencing reagent pins 172. The cleaning agent zone 18 can be divided as required into several cleaning agent reservoirs which are independent of one another, the separation contour line not being shown in the drawing.
In some embodiments, as shown in fig. 9, the purge flow path includes a second reversing valve assembly in communication with the common outlet of the second valve block 19, the tubing between the second reversing valve T5 and the ports B of the first through fourth pumping components P1-P4, and the outer walls of the plurality of second sequencing reagent pins 172, respectively, to selectively communicate the purge from the common outlet of the second valve block 19 with one of the tubing between the second reversing valve T5 and the ports B of the first through fourth pumping components P1-P4 and the outer walls of the plurality of second sequencing reagent pins 172.
In some embodiments, as shown in fig. 9, the second reversing valve assembly includes a third reversing valve T7, a fourth reversing valve T8, and a pump 21, the common outlet of the second valve assembly 19 is in communication with the second port of the fourth reversing valve T8, the third port of the fourth reversing valve T8 is in communication with the inlet of the pump 21, the outlet of the pump 21 is in communication with the second port of the third reversing valve T7, the third port of the third reversing valve T7 is in communication with the outer walls of the plurality of second sequencing reagent pins 172, the first ports of the third reversing valve T7 and the fourth reversing valve T8, and the line between the first port of the third reversing valve T7 and the first port of the fourth reversing valve T8 is in selective communication with the line between the second reversing valve T5 and the B ports of the first through fourth pumping components P1-P4 via a cut-off valve T6. Optionally, a one-to-many block 22 is provided in the line between the third port of the third reversing valve T7 and the outer walls of the plurality of second sequencing reagent pins 172, so that the cleaning agent from the third port of the third reversing valve T7 is split through the block 22 to the outer wall of the selected second sequencing reagent pin 172 for cleaning thereof. Alternatively, pump 21 may be adjustable in speed and counter-rotating, including, for example, diaphragm pumps and peristaltic pumps.
In some embodiments, as shown in fig. 9, the second valve block 19 further comprises another two-way valve T14, the inlet of the two-way valve T14 being in communication with the waste liquid zone 16, the outlet of the two-way valve T14 being in communication with the common outlet of the second valve block 19;
the purge flow path may perform at least one of the following operational steps:
step a: one of the two-way valves is selectively closed, and a corresponding cleaning agent is selected, which can be used to empty the liquid in the pipeline if the cleaning agent is air.
Step b: as shown in fig. 9, the third reversing valve T7 and the fourth reversing valve T8 are de-energized, and the pump 21 rotates forward to convey the selected cleaning agent from the cleaning agent zone 18 to the outer wall of the second sequencing reagent needle 172 of the sequencing reagent zone 17 via the second valve group 19, the fourth reversing valve T8, the pump 21, and the third reversing valve T7, and clean the outer wall of the second sequencing reagent needle 172.
Step c: as shown in fig. 10, the third reversing valve T7 is powered and the fourth reversing valve T8 is powered off, and the pump 21 is rotated in the forward direction so that the desired cleaning agent is drawn through the second valve block 19, the fourth reversing valve T8, the pump 21, the third reversing valve T7, and to the shut-off valve T6.
Step d: as shown in fig. 10, the cut-off valve T6 is turned on, the four first reversing valves T1-T4 are powered on and the second reversing valve T5 is powered off, and the selected cleaning agent sequentially passes through the cut-off valve T6, the second reversing valve T5, the fifth pipeline L5, the rotary valve 15, the second sequencing reagent needle 172, and finally enters the sequencing reagent tank of the sequencing reagent zone 17.
Step e: the rotary valve 15 is sequentially switched to each hole site, and step d is repeated to complete the cleaning of the inner wall of each second sequencing reagent needle 172 and the communication lines thereof.
F, executing the step a for multiple times to select different cleaning agents respectively, executing the step e, and cleaning the inner wall of each second sequencing reagent needle 172 and the communication pipeline thereof for multiple times by using the different cleaning agents;
step g: as shown in fig. 11, the third reversing valve T7 is powered on and the fourth reversing valve T8 is powered off, the four first reversing valves T1-T4 are powered on and the second reversing valve T5 is powered off, the pump 21 is rotated in reverse, and only the other two-way valve T14 of the second valve group is turned on, and the waste liquid in the sequencing reagent tank of the sequencing reagent section 17 is sucked into the waste liquid section 16.
And h, as shown in fig. 12, the stop valve T6 is connected, the reversing valves T1-4 are powered on, the second reversing valve T5 is powered off, the rotary valve 15 is switched to a plugged hole position, the ports B and O of the first to fourth pumping parts P1-P4 are simultaneously opened, the cleaning agent output in the step c sequentially passes through the quarter liquid block, the ports B and O of the first to fourth pumping parts P1-P4, and finally the waste liquid area 16 is discharged, so that the cleaning of related pipelines and valves is completed.
Step i: as shown in fig. 13, the third reversing valve T7 is de-energized and the fourth reversing valve T8 is energized, the pump 21 is turned off, and the selected cleaning agent passes through the outlet of the second valve block 19, the fourth reversing valve T8, and reaches the shut-off valve T6;
step j: as shown in fig. 13, the stop valve T6 is turned on, the first to second reversing valves T1-T5 are de-energized, the rotary valve 15 is switched to a hole site designated to be blocked, the first to fourth pumping parts P1-P4 are switched to an I port, a negative pressure pumping cleaning agent is provided, and the cleaning agent sequentially passes through the stop valve T6, the second reversing valve T5, the four biological channels C1-C4 of the test slide 13, the four first reversing valves T1-T4, the I ports of the first to fourth pumping parts P1-P4, and finally enters the inside of the first to fourth pumping parts P1-P4, thereby completing the cleaning of the relevant pipelines and valves.
Step k: the first to fourth pumping sections P1 to P4 are switched to the O-port, and positive pressure is supplied to discharge the cleaning agent therein into the waste liquid region 16 through the fourth line L4.
In the step l, as shown in fig. 14, the stop valve T6 is turned on, the four first reversing valves T1-T4 are powered on, the second reversing valve T5 is powered off, the rotary valve 15 is switched to a hole site appointed to be blocked, the first to fourth suction components P1-P4 are switched to the port B, negative pressure suction cleaning agent is provided, and the negative pressure suction cleaning agent sequentially passes through the stop valve T6, the quarter liquid block 23 and the port B of the first to fourth suction components P1-P4 and finally enters the first to fourth suction components P1-P4 for temporary storage.
In the step m, as shown in fig. 15, the four first reversing valves T1-T4 are powered on, the second reversing valve T5 is powered off, the first to fourth pumping parts P1-P4 are switched to an I port, positive pressure is provided to push the cleaning agent, the cleaning agent temporarily stored in the first to fourth pumping parts P1-P4 in the steps j and I is pushed out, the I port of the first to fourth pumping parts P1-P4 and the four first reversing valves T1-T4 are sequentially discharged into the first to fourth sample storage bins in the sample storage area 11 through four sample needles, and the cleaning of related pipelines and valves is completed.
Step n: as shown in fig. 16, the rotary valve 15 is switched to a hole position communicated with the fifth reversing valve T9, the four first reversing valves T1-4 are powered on, the second reversing valve T5 is powered off, the fifth reversing valve T9 is powered on, the first to fourth pumping parts P1-P4 are switched to a port B, positive pressure is provided to push out the cleaning agent temporarily stored in the first to fourth pumping parts P1-P4 in the j or l step, and the cleaning agent sequentially passes through the port B of the first to fourth pumping parts P1-P4, a quarter liquid block, the second reversing valve T5, the rotary valve 15, the fifth reversing valve T9, and finally is discharged to a fourth sample storage bin through a sample needle, thereby completing the cleaning of the related pipelines and valves.
According to the cleaning flow path, a plurality of different cleaning agents can be used for cleaning flow path components in the flow path system respectively, so that different cleaning requirements of the components are met, the cleaning efficiency is improved, the labor is saved, and the cleaning effect is better.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or equivalent substitutions can be made to some technical features thereof; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (10)
1. A method of loading a sequencing fluid onto a genetic sequencing slide, the genetic sequencing slide comprising a plurality of sample channels, the method comprising independently loading the plurality of sample channels with the sequencing fluid, the sequencing fluid comprising a biological sample, a sequencing reagent, or a cleaner.
2. The method of claim 1, wherein the step of loading the sequencing liquid comprises selectively flowing the sequencing liquid into one of the ends of the sample channel and out of the other of the ends of the sample channel such that the sequencing liquid can flow into one of the ends of the sample channel and also into the other of the ends of the sample channel.
3. The method of claim 2, wherein the step of loading a sequencing fluid comprises flowing the sequencing fluid through the sample channel by positive or negative pressure.
4. A flow path system for gene sequencing, comprising:
a genetic sequencing slide having a plurality of sample channels;
at least one of the following:
a biological sample loading flow path configured to be able to independently load a biological sample to the plurality of sample channels;
a sequencing reagent loading flow path configured to be capable of independently loading sequencing reagents into a plurality of sample channels; and
and a washing channel configured to be able to independently load washing agents to the plurality of sample channels.
5. The flow path system of claim 4, comprising at least one of:
a biological sample loading flow path configured to enable a biological sample to flow into the sample channel from one of both ends of the sample channel and to flow out from the other of both ends of the sample channel, so that the biological sample can flow into the sample channel from one end of the sample channel and also can flow into the sample channel from the other end of the sample channel;
a sequencing reagent loading flow path configured to enable a sequencing reagent to selectively flow into the sample channel from one of the two ends of the sample channel and to flow out of the other of the two ends of the sample channel, such that the sequencing reagent can flow into the sample channel from one end of the sample channel and also can flow into the sample channel from the other end of the sample channel; and
a purge flow path configured to enable a purge to flow into the sample channel from one of both ends of the sample channel selectively, and to flow out from the other of both ends of the sample channel, so that the purge can flow into the sample channel from one end of the sample channel or into the sample channel from the other end of the sample channel.
6. The flow path system of claim 5, comprising at least one of:
a biological sample loading flow path configured to flow a biological sample through the sample channel by positive or negative pressure;
a sequencing reagent loading flow path configured to flow a sequencing reagent through the sample channel by positive or negative pressure; and
a purge flow path configured to flow a purge through the sample channel by positive or negative pressure.
7. The flow path system of claim 4, wherein the flow path system comprises a plurality of gene sequencing slides that operate independently of one another.
8. The flow path system of claim 4, wherein one end of each of the plurality of sample channels in one of the gene sequencing slide is fluidly focused to a port such that sequencing fluid can flow into or out of the sample channels through the port.
9. A genetic sequencer, characterized in that the method according to any one of claims 1-3 is used.
10. A genetic sequencer comprising the flow system of any one of claims 4-8.
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| CNPCT/CN2023/105922 | 2023-07-05 |
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| CN202410298170.6A Pending CN117947141A (en) | 2023-07-05 | 2024-03-14 | Method for loading liquid for sequencing onto gene sequencing slide, flow path system for gene sequencing and gene sequencer |
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| CN117947141A (en) | 2024-04-30 |
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