WO2024065665A1 - 基因测序芯片、载片及其加工方法以及基因测序方法 - Google Patents

基因测序芯片、载片及其加工方法以及基因测序方法 Download PDF

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Publication number
WO2024065665A1
WO2024065665A1 PCT/CN2022/123237 CN2022123237W WO2024065665A1 WO 2024065665 A1 WO2024065665 A1 WO 2024065665A1 CN 2022123237 W CN2022123237 W CN 2022123237W WO 2024065665 A1 WO2024065665 A1 WO 2024065665A1
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Prior art keywords
gene
liquid outlet
gene sequencing
guide groove
sequencing chip
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English (en)
French (fr)
Inventor
卢剑
田畅
陆灏
王忠海
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MGI Tech Co Ltd
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MGI Tech Co Ltd
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Priority to CN202280096327.6A priority Critical patent/CN119325503A/zh
Priority to PCT/CN2022/123237 priority patent/WO2024065665A1/zh
Publication of WO2024065665A1 publication Critical patent/WO2024065665A1/zh
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • 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
    • C12M1/00—Apparatus for enzymology or microbiology
    • C12M1/34—Measuring or testing with condition measuring or sensing means, e.g. colony counters
    • 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

Definitions

  • the present application relates to the field of gene sequencing chip processing and use, and specifically, to a carrier, a gene sequencing chip, a method for processing a carrier of a gene sequencing chip, and a gene sequencing method.
  • the gene sequencing chip of a traditional high-throughput sequencer is a universal consumable that needs to be used in the corresponding gene sequencer.
  • the gene sequencing chip generally includes a carrier, a light-transmitting plate covering the carrier, and a frame structure arranged on the outer side of the light-transmitting plate and the carrier.
  • a reaction channel can be formed between the light-transmitting plate and the carrier for the flow of DNA molecular samples and reaction reagents.
  • the reaction channel of the current gene sequencing chip is relatively large, so the amount of DNA molecular samples and chemical reaction reagents required is large, and the cost of the reaction reagents is very high, which increases the cost of gene sequencing.
  • a large amount of DNA molecular samples and chemical reaction reagents will increase the test time, which is not conducive to rapid determination. If the size of the gene sequencing chip is changed, it will become difficult to use it with the gene sequencer.
  • the main purpose of the present application is to provide a carrier, a gene sequencing chip, a method for processing a carrier of a gene sequencing chip, and a gene sequencing method, so as to solve the problems of high cost and long testing time of gene sequencing chips in related technologies.
  • a carrier of a gene sequencing chip comprising: a substrate, comprising a gene attachment area and a gene barrier area, wherein a plurality of gene attachment points are arranged in the gene attachment area; a barrier coating, filled in the gene barrier area; a liquid inlet and a liquid outlet, both of which are arranged on the substrate, and both of which are connected to the gene attachment area.
  • a plurality of support columns are arranged in the gene attachment region, and a plurality of gene attachment points are arranged in the region outside the plurality of support columns.
  • the substrate is a polygonal structure
  • the substrate includes a first edge and a second edge that are intersectingly arranged
  • the gene attachment area is a proportionally reduced structure of the polygonal structure
  • the gene attachment area has a first boundary and a second boundary that intersect, the first edge coincides with the first boundary, the second edge coincides with the second boundary, the liquid inlet is arranged in the gene attachment area, the liquid outlet is arranged in the gene isolation area
  • the carrier also includes a guide groove arranged on the substrate, and the liquid inlet and the liquid outlet are connected through the guide groove.
  • the liquid outlet hole includes a first liquid outlet hole
  • the guide groove includes a first guide groove connecting the liquid inlet hole and the first liquid outlet hole
  • the liquid inlet hole is arranged at the first end of the first edge of the substrate
  • the first liquid outlet hole is arranged at the second end of the first edge of the substrate
  • the first guide groove is arranged along the first edge.
  • the substrate includes a third edge parallel to the first edge, the third edge includes a first end close to the second edge and a second end away from the second edge, the liquid outlet also includes a second liquid outlet arranged at the second end of the third edge, the guide groove also includes a second guide groove, the second guide groove includes a first groove section arranged on the gene attachment area and a second groove section arranged in the gene barrier area, the gene attachment area also has a third boundary parallel to the first boundary, the first groove section is arranged along the third boundary, and the second groove section is a straight groove section connected between an end of the first groove section away from the second boundary and the second liquid outlet.
  • the substrate is a first rectangular structure
  • the gene attachment region is a second rectangular structure
  • the ratio of the area of the gene attachment region to the area of the substrate is between 1/16 and 1/2.
  • the cross-section of the guide groove is a rectangular structure.
  • the cross section of the guide groove is a semicircular structure.
  • a gene sequencing chip comprising: a carrier, which is the carrier mentioned above; a light-transmitting plate, which covers the side of the carrier having the gene attachment area and the gene barrier area; a frame structure, which is arranged in the circumferential direction of the carrier and the light-transmitting plate, and the frame structure comprises a first mounting frame located above the light-transmitting plate and a second mounting frame located below the carrier, and the second mounting frame is provided with flow holes corresponding to the liquid inlet and outlet of the carrier.
  • a gene sequencing chip includes: a carrier, the carrier is the above-mentioned carrier; a light-transmitting plate, covering the side of the carrier having the gene attachment area and the gene barrier area; a frame structure, arranged in the circumferential direction of the carrier and the light-transmitting plate, the frame structure includes a first mounting frame arranged close to the light-transmitting plate and a second mounting frame close to the side of the carrier away from the light-transmitting plate, the carrier and the light-transmitting plate are clamped between the first mounting frame and the second mounting frame, and the second mounting frame is provided with a first flow hole corresponding to the liquid inlet hole of the carrier, a second flow hole corresponding to the first liquid outlet hole of the carrier, and a third flow hole corresponding to the second liquid outlet hole of the carrier.
  • a method for processing a carrier of a gene sequencing chip comprising: dividing a substrate into a first area and a second area, coating the first area with a barrier coating to form a gene barrier area, and forming a gene attachment area in the second area not coated with the barrier coating; and processing a liquid inlet hole and a liquid outlet hole on the substrate to communicate with the gene attachment area.
  • a gene sequencing method using the above-mentioned gene sequencing chip, the gene sequencing method comprising: contacting the gene sequencing chip with a plurality of reaction reagents in sequence according to a predetermined order, and cleaning the gene sequencing chip with a cleaning agent after each contact of the gene sequencing chip with a reaction reagent.
  • the step of making the gene sequencing chip contact with multiple reaction reagents in sequence according to a predetermined order includes: for each reaction reagent, when the gene sequencing chip contacts the reaction reagent, first making the reaction reagent flow along the first guide groove of the gene sequencing chip, and then making the reaction reagent flow along the second guide groove of the gene sequencing chip.
  • the steps of first allowing the reaction reagent to flow along the first guide groove of the gene sequencing chip and then allowing the reaction reagent to flow along the second guide groove of the gene sequencing chip include: opening the liquid inlet and the first liquid outlet, and introducing liquid through the liquid inlet to allow the reaction reagent to flow along the first guide groove of the gene sequencing chip; closing the first liquid outlet and opening the second liquid outlet to allow the reaction reagent to flow along the second guide groove of the gene sequencing chip.
  • the substrate includes a gene attachment area and a gene blocking area.
  • a plurality of gene attachment points are arranged in the gene attachment area for attaching DNA molecular samples.
  • a barrier coating is coated on the gene blocking area.
  • the gene attachment area can use the original liquid inlet and outlet holes on the substrate to realize the circulation of DNA molecular samples and reaction reagents.
  • the above-mentioned design method can reduce the reaction channel for the circulation of DNA molecular samples and reaction reagents without changing the size of the slide, thereby reducing the amount of reaction reagents, saving the cost of gene sequencing, and effectively shortening the sequencing time.
  • FIG1 shows a schematic structural diagram of a slide according to a first embodiment of the present application
  • Fig. 2 shows a cross-sectional view of the slide of Fig. 1 taken along line A-A;
  • FIG3 shows a cross-sectional view of a second embodiment of a slide according to the present application.
  • FIG. 4 shows a schematic structural diagram of a third embodiment of a slide according to the present application.
  • the carrier of the gene sequencing chip of the first embodiment includes: a substrate 10, a barrier coating 20, a liquid inlet 30 and a liquid outlet 40.
  • the substrate 10 includes a gene attachment area 11 and a gene barrier area 12, and a plurality of gene attachment points are arranged in the gene attachment area 11;
  • the barrier coating 20 is filled in the gene barrier area 12;
  • the liquid inlet 30 and the liquid outlet 40 are both arranged on the substrate 10, and the liquid inlet 30 and the liquid outlet 40 are both connected to the gene attachment area 11.
  • the substrate 10 includes a gene attachment area 11 and a gene isolation area 12.
  • a plurality of gene attachment points are arranged in the gene attachment area 11 for attaching DNA molecular samples.
  • the gene isolation area 12 is coated with a barrier coating 20.
  • the light-transmitting plate is covered on the substrate 10, an area for the circulation of DNA molecular samples and reaction reagents is formed between the gene attachment area 11 and the light-transmitting plate. Since the gene isolation area 12 is surrounded by the barrier coating 20, there is no way for DNA molecular samples and reaction reagents to enter between the gene isolation area 12 and the light-transmitting plate. Therefore, DNA molecular samples and reaction reagents can only enter the gene attachment area 11.
  • the gene attachment area 11 can use the original liquid inlet 30 and liquid outlet 40 on the substrate 10 to realize the circulation of DNA molecular samples and reaction reagents.
  • the above-mentioned design method can reduce the reaction channel for the circulation of DNA molecular samples and reaction reagents without changing the size of the slide, thereby reducing the amount of DNA molecular samples and reaction reagents, saving the cost of gene sequencing, and effectively shortening the reaction time.
  • the gene sequencing chip of the present application does not need to collect more samples to save reagents.
  • the gene sequencing chip of the present application can be compatible with the original structure of the sequencer without changing it.
  • carrier coating 20 filled in the gene isolation zone 12 may include the barrier coating completely filling the gene isolation zone 12, or may include the barrier coating 20 only filling the edge adjacent to the gene isolation zone 12 and the gene attachment zone 11 to prevent the DNA molecules and reaction reagents entering the gene attachment zone 11 from flowing into the gene isolation zone 12.
  • the present application reduces the amount of reagents used by changing the space for accommodating reagents, without changing the size of the substrate and thus without changing the relevant structure of the sequencer, thereby achieving a wide compatibility range at a minimum cost.
  • a plurality of support columns 13 are provided in the gene attachment area 11, and a plurality of gene attachment points are provided in the area outside the plurality of support columns 13.
  • the plurality of support columns 13 can support the light-transmitting plate, so that a reaction channel for the circulation of DNA molecular samples and reaction reagents is formed between the gene attachment area 11 and the light-transmitting plate, thereby facilitating the entry of DNA molecular samples and reaction reagents into the gene chip.
  • the support column 13 is made of the same material as the barrier coating 20, with one end bonded to the substrate 10 and the other end bonded to the light-transmitting plate, playing a supporting role on the one hand and a packaging role on the other.
  • the substrate 10 is a polygonal structure
  • the substrate 10 includes a first edge 14 and a second edge 15 that are intersectingly arranged
  • the gene attachment area 11 is a proportionally reduced structure of the polygonal structure
  • the gene attachment area 11 has a first boundary 111 and a second boundary 112 that are intersecting
  • the first edge 14 coincides with the first boundary 111
  • the second edge 15 coincides with the second boundary 112
  • the liquid inlet 30 is arranged in the gene attachment area 11
  • the liquid outlet 40 is arranged in the gene isolation area 12
  • the slide also includes a guide groove 50 arranged on the substrate 10, and the liquid inlet 30 and the liquid outlet 40 are connected through the guide groove 50.
  • the gene attachment area 11 is made to coincide with the edge of the substrate 10 so that the original structure on the substrate can be applied as much as possible, thereby simplifying the modification process of the substrate 10 and facilitating the processing of the substrate 10.
  • the gene attachment area can also be set in the central area of the substrate, and the gene attachment area is connected to the original liquid inlet and outlet holes of the substrate by opening the guide groove.
  • the liquid outlet hole 40 includes a first liquid outlet hole 41
  • the guide groove 50 includes a first guide groove 51 connecting the liquid inlet hole 30 and the first liquid outlet hole 41
  • the liquid inlet hole 30 is arranged at the first end of the first edge 14 of the substrate 10
  • the first liquid outlet hole 41 is arranged at the second end of the first edge 14 of the substrate 10
  • the first guide groove 51 is arranged along the first edge 14.
  • liquid inlet 30 is located in the gene attachment area 11 and is directly connected to the reaction channel for the entry of DNA molecule samples and reaction reagents.
  • the first liquid outlet 41 is located in the gene isolation area 12.
  • the barrier coating 20 needs to avoid the first guide groove 51 located in the gene isolation area 12 so that the first guide groove 51 is connected to the reaction channel.
  • the substrate 10 includes a third edge 16 parallel to the first edge 14, the third edge 16 includes a first end close to the second edge 15 and a second end away from the second edge 15, the liquid outlet 40 also includes a second liquid outlet 42 arranged at the second end of the third edge 16,
  • the guide groove 50 also includes a second guide groove 52, the second guide groove 52 includes a first groove section 521 arranged on the gene attachment area 11 and a second groove section 522 arranged in the gene isolation area 12, the gene attachment area 11 also has a third boundary 113 parallel to the first boundary 111, the first groove section 521 is arranged along the third boundary 113, and the second groove section 522 is a straight groove section connected between the end of the first groove section 521 away from the second boundary 112 and the second liquid outlet 42.
  • the slide of this embodiment includes two guide grooves, namely the first guide groove 51 and the second guide groove 52. Specifically, in actual operation, the DNA molecule sample or the reaction reagent can first flow along the first guide groove 51 of the gene sequencing chip, and then the reaction reagent can flow along the second guide groove 52 of the gene sequencing chip.
  • the specific steps include: opening the liquid inlet 30 and the first liquid outlet 41, and feeding liquid through the liquid inlet 30, so that the DNA molecule sample or the reaction reagent flows along the first guide groove 51 of the gene sequencing chip; closing the first liquid outlet 41, and opening the second liquid outlet 42, so that the reaction reagent flows through the gene attachment area 11 and then flows along the second guide groove 52 of the gene sequencing chip.
  • the DNA molecule sample or the reaction reagent can be quickly flowed into the reaction channel by positive pressure injection at the liquid inlet 30, or by negative pressure at the first liquid outlet 41 or the second liquid outlet 42.
  • the substrate 10 is a first rectangular structure
  • the gene attachment area 11 is a second rectangular structure
  • the ratio of the area of the gene attachment area 11 to the area of the substrate 10 is between 1/16 and 1/2.
  • the area of the above-mentioned gene attachment area 11 and the area of the substrate 10 can be flexibly designed according to actual reaction requirements.
  • the ratio of the area of the gene attachment area 11 to the area of the substrate 10 can be 1/16, 1/10, 1/8, 1/6, 1/4 or 1/2.
  • the ratio of the area of the gene attachment area 11 to the area of the substrate 10 is 1/4.
  • the cross section of the flow guide groove 50 is a semicircular structure, and its radius is generally one order of magnitude larger than the height of the reaction channel.
  • the above structure makes the bottom wall of the flow guide groove 50 smoother, so that the reaction reagent is not easy to remain in the flow guide groove 50, thereby reducing the risk of mixing the reaction reagent that enters the gene chip later with the reaction reagent that enters the gene chip earlier, and reducing the probability of the reaction reagent being contaminated.
  • the present application also provides a second embodiment of the slide, and the difference between the second embodiment and the first embodiment is that the structure of the flow guide groove 50 is different.
  • the cross section of the flow guide groove 50 is a rectangular structure, and its depth and width are generally one order of magnitude greater than the height of the reaction channel. The above structure facilitates the processing of the flow guide groove 50, further reducing the difficulty of improving the slide.
  • the present application also provides a third embodiment of the slide, and the difference between the third embodiment and the first and second embodiments is that the specific shape of the gene attachment area 11 is different.
  • the third embodiment the three edges of the gene attachment area 11 overlap with the three edges of the substrate 10, and the ratio of its area to the area of the substrate 10 is between 1/16 and 1/4.
  • the above-mentioned setting makes the width of the gene attachment area 11 smaller, and only the first guide groove 51 is used to make the DNA molecule sample and the reaction reagent evenly distributed in the gene attachment area 11.
  • the shape of the gene attachment area in the third embodiment can be designed as a trapezoid, and the liquid inlet and outlet are respectively located at the two bottom corners of the trapezoid, which is conducive to more fully replacing the reagent and reducing the dead volume.
  • the guide groove in the fourth embodiment can be designed to be parallel to one of the waists of the trapezoid.
  • the shape of the gene attachment area, the number, position and shape of the guide grooves, and the number and position of the liquid inlet and outlet holes can all be flexibly designed according to actual conditions.
  • the present application also provides a gene sequencing chip
  • an embodiment of the gene sequencing chip of the present application includes: a carrier, a light-transmitting plate, and a frame structure.
  • the carrier is the above-mentioned carrier
  • the light-transmitting plate is covered on one side of the carrier having the gene attachment area 11 and the gene blocking area 12
  • the frame structure is arranged in the circumferential direction of the carrier and the light-transmitting plate, and the frame structure includes a first mounting frame located above the light-transmitting plate and a second mounting frame located below the carrier, and the second mounting frame is provided with flow holes corresponding to the liquid inlet hole 30 and the liquid outlet hole 40 of the carrier.
  • the carrier since the carrier has the advantages of reducing the cost of gene sequencing, fast reaction, and small application amount of DNA molecular samples, the gene sequencing chip having it also has the above advantages.
  • the present application also provides a gene sequencing chip
  • the embodiment of the gene sequencing chip of the present application includes: a carrier, a light-transmitting plate, and a frame structure.
  • the carrier is the above-mentioned carrier
  • the light-transmitting plate is covered on the side of the carrier having the gene attachment area 11 and the gene blocking area 12
  • the frame structure is arranged in the circumferential direction of the carrier and the light-transmitting plate, and the frame structure includes a first mounting frame arranged close to the light-transmitting plate and a second mounting frame arranged close to the side of the carrier away from the light-transmitting plate, the carrier and the light-transmitting plate are sandwiched between the first mounting frame and the second mounting frame
  • the second mounting frame is provided with a first flow hole arranged corresponding to the liquid inlet 30 of the carrier, a second flow hole arranged corresponding to the first liquid outlet 41 of the carrier, and a third flow hole arranged corresponding to the second liquid outlet 42 of
  • the above structure enables the gene sequencing chip to be used in conjunction with a gene sequencer, ensuring the versatility of the gene sequencing chip.
  • the carrier since the carrier has the advantages of reducing the cost of gene sequencing, fast reaction, and small amount of reagent application, the gene sequencing chip having it also has the above advantages.
  • the present application also provides a method for processing a slide of a gene sequencing chip.
  • the embodiment of the method for processing a slide of a gene sequencing chip includes: dividing a substrate 10 into a first area and a second area, coating the first area with a barrier coating 20 to form a gene barrier area 12, and forming a gene attachment area 11 in the second area not surrounded by the barrier coating 20; processing a liquid inlet 30 and a liquid outlet 40 connected to the gene attachment area 11 on the substrate 10.
  • the present application also provides a gene sequencing method, using the above-mentioned gene sequencing chip, the gene sequencing method comprising: contacting the gene sequencing chip with a plurality of reaction reagents in sequence according to a predetermined order, and using a cleaning agent to clean the gene sequencing chip after each contact of the gene sequencing chip with a reaction reagent.
  • the above-mentioned structure can make it difficult for the reaction reagent to remain in the gene sequencing chip, thereby reducing the risk of mixing the reaction reagent that enters the gene chip later with the reaction reagent that enters the gene chip earlier, and reducing the probability of the reaction reagent being contaminated.
  • the step of making the gene sequencing chip contact with multiple reaction reagents in a predetermined order includes: for each reaction reagent, when the gene sequencing chip contacts the reaction reagent, the reaction reagent is first made to flow along the first flow guide groove 51 of the gene sequencing chip, and then the reaction reagent is made to flow along the second flow guide groove 52 of the gene sequencing chip.
  • the above structure can ensure that the reaction reagent can be evenly diffused and distributed in the gene attachment area 11, thereby achieving a better gene sequencing effect and making full use of the reagents.
  • the steps of first making the reaction reagent flow along the first flow guide groove 51 of the gene sequencing chip, and then making the reaction reagent flow along the second flow guide groove 52 of the gene sequencing chip include: opening the liquid inlet and the first liquid outlet, and feeding liquid through the liquid inlet to make the reaction reagent flow along the first flow guide groove 51 of the gene sequencing chip; closing the first liquid outlet, and opening the second liquid outlet to make the reaction reagent diffuse and flow along the second flow guide groove 52 of the gene sequencing chip.
  • the above structure can further ensure that the reaction reagent can be evenly diffused and distributed in the gene attachment area 11. This facilitates subsequent fluorescent labeling and fluorescent imaging.

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Abstract

本申请提供了一种基因测序芯片的载片、基因测序芯片、基因测序芯片的载片的加工方法以及基因测序方法。其中，基因测序芯片的载片，包括：基板，包括基因附着区和基因阻隔区，基因附着区内设置有多个基因附着点位；阻隔涂层，涂布在基因阻隔区的轮廓边缘或者填充在基因阻隔区内；进液孔和出液孔，均设置于基板上，进液孔和出液孔均与基因附着区连通。应用本申请的技术方案通过更改容纳试剂的空间以减少试剂用量，不需要更改基板的尺寸，进而不需要更改测序仪的有关构造，以最小的代价实现宽泛的兼容范围。

Description

基因测序芯片、载片及其加工方法以及基因测序方法 技术领域
本申请涉及基因测序芯片加工和使用领域，具体而言，涉及一种载片、基因测序芯片、基因测序芯片的载片的加工方法、基因测序方法。
背景技术
随基因测序技术的发展，通过阵列式反应池基因测序芯片进行荧光标记、荧光成像、去阻断、去除荧光基团等连续循环的化学反应的测序技术目前已广泛应用于全基因组测序，转录组测序，宏基因组测序等试验中。相应地，容纳基因测序芯片的基因测序平台和产品不断更迭，基因测序芯片作为特定DNA片段的碱基序列测序过程的重要器件，在测序技术的推动下正朝高通量、低成本方向发展。
传统高通量测序仪的基因测序芯片是一种通用耗材，需要应用在对应的基因测序仪上，基因测序芯片一般包括载片，盖在载片上的透光板以及设置在透光板和载片周向外侧的框架结构，透光板和载片之间能够形成供DNA分子样本和反应试剂流通的反应通道。
目前的基因测序芯片的反应通道的体积较大，因此所需的DNA分子样本和化学反应试剂的用量大，反应试剂的成本很高，使得基因测序的成本提高，并且大量的DNA分子样本和化学反应试剂会使得测试时间增长，不利于快速测定。而如果改变基因测序芯片的尺寸则会导致其与基因测序仪难以配合使用。
申请内容
本申请的主要目的在于提供一种载片、基因测序芯片、基因测序芯片的载片的加工方法、基因测序方法，以解决相关技术中的基因测序芯片测试成本高并且测试时间长的问题。
为了实现上述目的，根据本申请的一个方面，提供了一种基因测序芯片的载片，包括：基板，包括基因附着区和基因阻隔区，基因附着区内设置有多个基因附着点位；阻隔涂层，填充在基因阻隔区内；进液孔和出液孔，均设置于基板上，进液孔和出液孔均与基因附着区连通。
进一步地，基因附着区内设置有多个支撑柱，多个基因附着点位设置在多个支撑柱以外的区域内。
进一步地，基板为多边形结构，基板包括相交设置的第一边沿和第二边沿，基因附着区为多边形结构的等比例缩小结构，基因附着区具有相交的第一边界和第二边界，第一边沿与第一边界重合，第二边沿和第二边界重合，进液孔设置于基因附着区内，出液孔设置于基因阻隔区内，载片还包括设置于基板上的导流槽，进液孔和出液孔通过导流槽连通。
进一步地，出液孔包括第一出液孔，导流槽包括连通进液孔和第一出液孔的第一导流槽，进液孔设置于基板的第一边沿的第一端处，第一出液孔设置于基板的第一边沿的第二端处，第一导流槽沿第一边沿设置。
进一步地，基板包括平行于第一边沿的第三边沿，第三边沿包括靠近第二边沿的第一端和远离第二边沿的第二端，出液孔还包括设置于第三边沿的第二端处的第二出液孔，导流槽还包括第二导流槽，第二导流槽包括设置于基因附着区上的第一槽段和设置于基因阻隔区内的第二槽段，基因附着区还具有平行于第一边界平行的第三边界，第一槽段沿第三边界设置，第二槽段为连通在第一槽段的远离第二边界的一端与第二出液孔之间的直线槽段。
进一步地，基板为第一矩形结构，基因附着区为第二矩形结构，基因附着区的面积与基板的面积的比值在1/16至1/2之间。
进一步地，导流槽的截面为矩形结构。
进一步地，导流槽的截面为半圆形结构。
根据本申请的另一方面，提供了一种基因测序芯片，包括：载片，载片为上述的载片；透光板，盖设于载片的具有基因附着区和基因阻隔区的一侧；框架结构，设置于载片和透光板的周向方向上，框架结构包括位于透光板上方的第一安装框和位于载片下方的第二安装框，第二安装框上设置有与载片的进液孔和出液孔对应设置过流孔。
根据本申请的另一方面，提供了一种基因测序芯片，其特征在于，包括：载片，载片为上述的载片；透光板，盖设于载片的具有基因附着区和基因阻隔区的一侧；框架结构，设置于载片和透光板的周向方向上，框架结构包括靠近透光板设置的第一安装框和靠近载片的远离透光板的一侧的第二安装框，载片和透光板夹设于第一安装框和第二安装框之间，第二安装框上设置有与载片的进液孔对应设置的第一过流孔、与载片的第一出液孔对应设置的第二过流孔以及与载片的第二出液孔对应设置的第三过流孔。
根据本申请的另一方面，提供了一种基因测序芯片的载片的加工方法，包括：在基板上划分形成第一区域和第二区域，在第一区域内涂布阻隔涂层以形成基因阻隔区， 未涂布阻隔涂层的第二区域形成基因附着区域；在基板上加工出与基因附着区连通的进液孔和出液孔。
根据本申请的另一方面，提供了一种基因测序方法，采用上述的基因测序芯片，基因测序方法包括：使基因测序芯片按照预定顺序依次与多种反应试剂接触，并在每次基因测序芯片与一种反应试剂接触后采用清洗剂对基因测序芯片进行清洗。
进一步地，使基因测序芯片按照预定顺序依次与多种反应试剂接触的步骤包括：针对每种反应试剂，在基因测序芯片与该反应试剂接触时，先使反应试剂沿基因测序芯片的第一导流槽流动，再使反应试剂沿基因测序芯片的第二导流槽流动。
进一步地，在基因测序芯片与一种反应试剂接触时，先使反应试剂沿基因测序芯片的第一导流槽流动，再使反应试剂沿基因测序芯片的第二导流槽流动的步骤包括：开启进液孔和第一出液孔，通过进液孔进行进液，以使反应试剂沿基因测序芯片的第一导流槽流动；关闭第一出液孔，开启第二出液孔，以使反应试剂沿基因测序芯片的第二导流槽流动。
应用本申请的技术方案，基板包括基因附着区和基因阻隔区，基因附着区内设置有多个基因附着点位，用于附着DNA分子样本。基因阻隔区上涂布有阻隔涂层，当与载片配合的透光板盖在载片的基板上时，基因附着区和透光板之间形成供DNA分子样本和反应试剂流通的区域，而基因阻隔区上因为具有阻隔涂层，基因阻隔区与透光板之间没有供DNA分子样本和反应试剂进入的途径，因此DNA分子样本和反应试剂仅能够进入基因附着区内。基因附着区可以采用基板上原有的进液孔和出液孔实现DNA分子样本和反应试剂的流通。上述设计方式能够在不改变载片尺寸的情况下，缩小供DNA分子样本和反应试剂流通的反应通道，从而降低了反应试剂的用量，节约了基因测序的成本，并且能够有效缩短测序时间。
附图说明
构成本申请的一部分的说明书附图用来提供对本申请的进一步理解，本申请的示意性实施例及其说明用于解释本申请，并不构成对本申请的不当限定。在附图中：
图1示出了根据本申请的载片的实施例一的结构示意图；
图2示出了图1的载片的A-A向的剖视图；
图3示出了根据本申请的载片的实施例二的剖视图；
图4示出了根据本申请的载片的实施例三的结构示意图。
其中，上述附图包括以下附图标记：
10、基板；11、基因附着区；111、第一边界；112、第二边界；113、第三边界；12、基因阻隔区；13、支撑柱；14、第一边沿；15、第二边沿；16、第三边沿；20、阻隔涂层；30、进液孔；40、出液孔；41、第一出液孔；42、第二出液孔；50、导流槽；51、第一导流槽；52、第二导流槽；521、第一槽段；522、第二槽段。
具体实施方式
下面将结合本申请实施例中的附图，对本申请实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本申请一部分实施例，而不是全部的实施例。以下对至少一个示例性实施例的描述实际上仅仅是说明性的，决不作为对本申请及其应用或使用的任何限制。基于本申请中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例，都属于本申请保护的范围。
如图1至图3所示，实施例一的基因测序芯片的载片包括：基板10、阻隔涂层20、进液孔30以及出液孔40。其中，基板10包括基因附着区11和基因阻隔区12，基因附着区11内设置有多个基因附着点位；阻隔涂层20填充在基因阻隔区12内；进液孔30和出液孔40均设置于基板10上，进液孔30和出液孔40均与基因附着区11连通。
应用本实施的技术方案，基板10包括基因附着区11和基因阻隔区12，基因附着区11内设置有多个基因附着点位，用于附着DNA分子样本。基因阻隔区12上涂布有阻隔涂层20，当透光板盖在基板10上时，基因附着区11和透光板之间形成供DNA分子样本和反应试剂流通的区域，而基因阻隔区12上因为具有阻隔涂层20围合，基因阻隔区12与透光板之间没有供DNA分子样本和反应试剂进入的途径，因此DNA分子样本和反应试剂仅能够进入基因附着区11内。基因附着区11可以采用基板10上原有的进液孔30和出液孔40实现DNA分子样本和反应试剂的流通。上述设计方式能够在不改变载片尺寸的情况下，缩小供DNA分子样本和反应试剂流通的反应通道，从而降低了DNA分子样本和反应试剂的用量，节约了基因测序的成本，并且能够有效缩短反应时间。此外，DNA样本的量不多时，采用本申请的基因测序芯片可不必凑集更多的样本以节约试剂。同时，由于芯片外轮廓尺寸不变，可在不更改测序仪原有构造的情况下兼容本申请的基因测序芯片。
需要说明的是，上述的“阻隔涂层20填充在基因阻隔区12内”可以包括阻隔涂层将基因阻隔区12完全填满，也可以包括阻隔涂层20仅填充在基因阻隔区12与基因附着区11相邻的边沿处，以防止进入基因附着区11内的DNA分子和反应试剂流入到基因阻隔区12内。
综合比较，本申请通过更改容纳试剂的空间以减少试剂用量，不需要更改基板的尺寸，进而不需要更改测序仪的有关构造，以最小的代价实现宽泛的兼容范围。
如图1至图3所示，在实施例一中，基因附着区11内设置有多个支撑柱13，多个基因附着点位设置在多个支撑柱13以外的区域内。上述结构中，当透光板盖在基板10上时，多个支撑柱13能够支撑透光板，使得基因附着区11与透光板之间形成供DNA分子样本和反应试剂流通的反应通道，从而便于DNA分子样本和反应试剂进入到基因芯片内。而基因阻隔区12内由于设置阻隔涂层20，当透光板盖在基板10上时，阻隔涂层20与透光板之间形成密闭空间或者紧贴无间隙，从而有效防止DNA分子样本和反应试剂进入。一般地，支撑柱13与阻隔涂层20材料相同，一端粘接基板10，另一端粘接透光板，一方面起支撑作用，另一方面起封装作用。
如图1至图3所示，在实施例一中，基板10为多边形结构，基板10包括相交设置的第一边沿14和第二边沿15，基因附着区11为多边形结构的等比例缩小结构，基因附着区11具有相交的第一边界111和第二边界112，第一边沿14与第一边界111重合，第二边沿15和第二边界112重合，进液孔30设置于基因附着区11内，出液孔40设置于基因阻隔区12内，载片还包括设置于基板10上的导流槽50，进液孔30和出液孔40通过导流槽50连通。上述结构中，由于基板上原有的进液孔、出液孔以及导流槽均设置在基板的边缘处，因此使基因附着区11与基板10的边沿重合能够尽可能地应用基板上原有的结构，从而简化基板10的改造工艺，便于基板10的加工。
当然，在图中未示出的其他实施例中，基因附着区也可以设置在基板的中心区域，通过开设导流槽使得基因附着区与基板原有的进液孔和出液孔连通。
如图1至图3所示，在本实施例中，出液孔40包括第一出液孔41，导流槽50包括连通进液孔30和第一出液孔41的第一导流槽51，进液孔30设置于基板10的第一边沿14的第一端处，第一出液孔41设置于基板10的第一边沿14的第二端处，第一导流槽51沿第一边沿14设置。上述机构利用了基板上原有的进液孔、出液孔以及导流槽，从而简化基板10的改造工艺，便于基板10的加工。
需要说明的是，进液孔30位于基因附着区11，与供DNA分子样本和反应试剂进入的反应通道直接连通，第一出液孔41位于基因阻隔区12内，阻隔涂层20需要避让位于基因阻隔区12内的第一导流槽51，以使第一导流槽51与反应通道连通。
如图1至图3所示，在实施例一中，基板10包括平行于第一边沿14的第三边沿16，第三边沿16包括靠近第二边沿15的第一端和远离第二边沿15的第二端，出液孔40还包括设置于第三边沿16的第二端处的第二出液孔42，导流槽50还包括第二导流槽52，第二导流槽52包括设置于基因附着区11上的第一槽段521和设置于基因阻隔区12内的第二槽段522，基因附着区11还具有平行于第一边界111平行的第三边界113，第一槽段521沿第三边界113设置，第二槽段522为连通在第一槽段521的远离第二边界112的一端与第二出液孔42之间的直线槽段。
上述机构中，为了便于后续的荧光标记以及荧光成像，需要使DNA分子样本以及反应试剂能够均匀地分布在基因附着区11内。因此本实施例的载片包括两条导流槽，即第一导流槽51和第二导流槽52，具体地，在实际操作中，可先使DNA分子样本或者反应试剂沿基因测序芯片的第一导流槽51流动，再使反应试剂沿基因测序芯片的第二导流槽52流动，具体步骤包括：开启进液孔30和第一出液孔41，通过进液孔30进行进液，以使DNA分子样本或者反应试剂沿基因测序芯片的第一导流槽51流动；关闭第一出液孔41，开启第二出液孔42，以使反应试剂流经基因附着区11后沿基因测序芯片的第二导流槽52流动。
需要说明的是，可以采用在进液孔30正压注液的方式使DNA分子样本或者反应试剂快速流入反应通道内，也可以采用在第一出液孔41或者第二出液孔42分别加负压的方式使得DNA分子样本或者反应试剂快速流入反应通道内。
具体地，如图1至图3所示，基板10为第一矩形结构，基因附着区11为第二矩形结构，基因附着区11的面积与基板10的面积的比值在1/16至1/2之间。上述基因附着区11的面积与基板10的面积可以根据实际的反应需求灵活设计。具体地，基因附着区11的面积与基板10的面积的比值可以为1/16、1/10、1/8、1/6、1/4或者1/2。在实施例一中，基因附着区11的面积与基板10的面积的比值为1/4。
如图2所示，在实施例一中，导流槽50的截面为半圆形结构，其半径尺寸一般比反应通道的高度大一个数量级。上述结构使得导流槽50的底壁更加平滑，从而使得反应试剂不易在导流槽50内残留，从而降低了后进入基因芯片内的反应试剂与先进入基因芯片内的反应试剂混合的风险，降低了反应试剂被污染的概率。
如图3所示，本申请还提供了载片的第二种实施例，实施例二与实施例一的区别在于导流槽50的结构不同。具体地，在实施例二中，导流槽50的截面为矩形结构，其深度和宽度一般比反应通道的高度大一个数量级。上述结构便于导流槽50的加工，进一步降低了载片的改进难度。
如图4所示，本申请还提供了载片的第三种实施例，实施例三与实施例一和实施例二的区别在于：基因附着区11的具体形状不同。具体地，在实施例三中，基因附着区11的三个边沿分别于基板10的三个边沿重合，其面积与基板10的面积比在1/16至1/4之间，上述设置方式使得基因附着区11的宽度较小，仅采用第一导流槽51即可使得DNA分子样本和反应试剂在基因附着区11内均匀分布。
另外，在图中未示出的第四种实施例中，可以将第三种实施例中基因附着区的形状设计为梯形，进液孔和出液孔分别位于梯形的两个底角处，有利于更充分替代试剂， 减少死体积。进一步地，第四种实施例中的导流槽可以设计为平行于梯形的其中一条腰。
当然，在其他实施例中，基因附着区的形状，导流槽的数量、位置及形状，进液孔和出液孔数量及位置，均可根据实际情况灵活设计。
根据本申请的另一方面，本申请还提供了一种基因测序芯片，本申请的基因测序芯片的实施例包括：载片、透光板、框架结构。其中，载片为上述的载片；透光板盖设于载片的具有基因附着区11和基因阻隔区12的一侧；框架结构设置于载片和透光板的周向方向上，框架结构包括位于透光板上方的第一安装框和位于载片下方的第二安装框，第二安装框上设置有与载片的进液孔30和出液孔40对应设置过流孔。上述结构中，由于载片具有降低基因测序成本、反应快、DNA分子样本应用量小的优点，因此具有其的基因测序芯片也具备上述优点。
根据本申请的另一方面，本申请还提供了一种基因测序芯片，本申请的基因测序芯片的实施例包括：载片、透光板、框架结构。其中，载片为上述的载片；透光板盖设于载片的具有基因附着区11和基因阻隔区12的一侧；框架结构设置于载片和透光板的周向方向上，框架结构包括靠近透光板设置的第一安装框和靠近载片的远离透光板的一侧的第二安装框，载片和透光板夹设于第一安装框和第二安装框之间，第二安装框上设置有与载片的进液孔30对应设置的第一过流孔、与载片的第一出液孔41对应设置的第二过流孔以及与载片的第二出液孔42对应设置的第三过流孔。上述结构使得基因测序芯片能够配合基因测序仪使用，保证了基因测序芯片的通用性。另外，由于载片具有降低基因测序成本、反应快、试剂应用量小的优点，因此具有其的基因测序芯片也具备上述优点。
根据本申请的另一方面，本申请还提供了一种基因测序芯片的载片的加工方法，基因测序芯片的载片的加工方法的实施例包括：在基板10上划分形成第一区域和第二区域，在第一区域内涂布阻隔涂层20以形成基因阻隔区12，未被阻隔涂层20围合的第二区域形成基因附着区11；在基板10上加工出与基因附着区11连通的进液孔30和出液孔40。上述步骤简单，操作方便，使得操作人员在原有载片的基础上进行简单改进即可获得本申请的载片。
根据本申请的另一方面，本申请还提供了一种基因测序方法，采用上述的基因测序芯片，基因测序方法包括：使基因测序芯片按照预定顺序依次与多种反应试剂接触，并在每次基因测序芯片与一种反应试剂接触后采用清洗剂对基因测序芯片进行清洗。上述结构能够使得反应试剂不易在基因测序芯片内残留，从而降低了后进入基因芯片内的反应试剂与先进入基因芯片内的反应试剂混合的风险，降低了反应试剂被污染的概率。
如图1所示，在本实施例中，使基因测序芯片按照预定顺序依次与多种反应试剂接触的步骤包括：针对每种反应试剂，在基因测序芯片与该反应试剂接触时，先使反应试剂沿基因测序芯片的第一导流槽51流动，再使反应试剂沿基因测序芯片的第二导流槽52流动。上述结构能够保证反应试剂在基因附着区11内能够均匀扩散分布，从而起到更好的基因测序效果，充分利用试剂。
如图1所示，在本实施例中，在基因测序芯片与一种反应试剂接触时，先使反应试剂沿基因测序芯片的第一导流槽51流动，再使反应试剂沿基因测序芯片的第二导流槽52流动的步骤包括：开启进液孔和第一出液孔，通过进液孔进行进液，以使反应试剂沿基因测序芯片的第一导流槽51流动；关闭第一出液孔，开启第二出液孔，以使反应试剂沿基因测序芯片的第二导流槽52扩散流动。上述结构能够进一步地保证反应试剂在基因附着区11内能够均匀扩散分布。从而便于后续的荧光标记以及荧光成像。
以上所述仅为本申请的优选实施例而已，并不用于限制本申请，对于本领域的技术人员来说，本申请可以有各种更改和变化。凡在本申请的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本申请的保护范围之内。

Claims (14)

  1. 一种基因测序芯片的载片，其特征在于，包括：
    基板(10)，包括基因附着区(11)和基因阻隔区(12)，所述基因附着区(11)内设置有多个基因附着点位；
    阻隔涂层(20)，填充在所述基因阻隔区(12)内；
    进液孔(30)和出液孔(40)，均设置于所述基板(10)上，所述进液孔(30)和所述出液孔(40)均与所述基因附着区(11)连通。
  2. 根据权利要求1所述的载片，其特征在于，所述基因附着区(11)内设置有多个支撑柱(13)，多个所述基因附着点位设置在多个所述支撑柱(13)以外的区域内。
  3. 根据权利要求1所述的载片，其特征在于，所述基板(10)为多边形结构，所述基板(10)包括相交设置的第一边沿(14)和第二边沿(15)，所述基因附着区(11)为所述多边形结构的等比例缩小结构，所述基因附着区(11)具有相交的第一边界(111)和第二边界(112)，所述第一边沿(14)与所述第一边界(111)重合，所述第二边沿(15)和所述第二边界(112)重合，所述进液孔(30)设置于所述基因附着区(11)内，所述出液孔(40)设置于所述基因阻隔区(12)内，所述载片还包括设置于所述基板(10)上的导流槽(50)，所述进液孔(30)和所述出液孔(40)通过所述导流槽(50)连通。
  4. 根据权利要求3所述的载片，其特征在于，所述出液孔(40)包括第一出液孔(41)，所述导流槽(50)包括连通所述进液孔(30)和所述第一出液孔(41)的第一导流槽(51)，所述进液孔(30)设置于所述基板(10)的第一边沿(14)的第一端处，所述第一出液孔(41)设置于所述基板(10)的第一边沿(14)的第二端处，所述第一导流槽(51)沿所述第一边沿(14)设置。
  5. 根据权利要求4所述的载片，其特征在于，所述基板(10)包括平行于所述第一边沿(14)的第三边沿(16)，所述第三边沿(16)包括靠近所述第二边沿(15)的第一端和远离所述第二边沿(15)的第二端，所述出液孔(40)还包括设置于所述第三边沿(16)的第二端处的第二出液孔(42)，所述导流槽(50)还包括第二导流槽(52)，所述第二导流槽(52)包括设置于所述基因附着区(11)上的第一槽段(521)和设置于所述基因阻隔区(12)内的第二槽段(522)，所述基因附着区(11)还具有平行于第一边界(111)平行的第三边界(113)，所述第一槽段(521)沿所述第三边界(113)设置，所述第二槽段(522)为连通在所述第一槽段(521)的远离所述第二边界(112)的一端与所述第二出液孔(42)之间的直线槽段。
  6. 根据权利要求1至5中任一项所述的载片，其特征在于，所述基板(10)为第一矩形结构，基因附着区(11)为第二矩形结构或为梯形结构，所述基因附着区(11)的面积与所述基板(10)的面积的比值在1/16至1/2之间。
  7. 根据权利要求3至5中任一项所述的载片，其特征在于，所述导流槽(50)的截面为矩形结构。
  8. 根据权利要求3至5中任一项所述的载片，其特征在于，所述导流槽(50)的截面为半圆形结构。
  9. 一种基因测序芯片，其特征在于，包括：
    载片，所述载片为权利要求1至8中任一项所述的载片；
    透光板，盖设于所述载片的具有所述基因附着区(11)和所述基因阻隔区(12)的一侧；
    框架结构，设置于所述载片和所述透光板的周向方向上，所述框架结构包括位于所述透光板上方的第一安装框和位于所述载片下方的第二安装框，所述第二安装框上设置有与所述载片的进液孔(30)和出液孔(40)对应设置过流孔。
  10. 一种基因测序芯片，其特征在于，包括：
    载片，所述载片为权利要求5所述的载片；
    透光板，盖设于所述载片的具有所述基因附着区(11)和所述基因阻隔区(12)的一侧；
    框架结构，设置于所述载片和所述透光板的周向方向上，所述框架结构包括靠近所述透光板设置的第一安装框和靠近所述载片的远离所述透光板的一侧的第二安装框，所述载片和所述透光板夹设于所述第一安装框和所述第二安装框之间，所述第二安装框上设置有与所述载片的进液孔(30)对应设置的第一过流孔、与所述载片的第一出液孔(41)对应设置的第二过流孔以及与所述载片的第二出液孔(42)对应设置的第三过流孔。
  11. 一种基因测序芯片的载片的加工方法，其特征在于，包括：
    在基板(10)上划分形成第一区域和第二区域，在第一区域内涂布阻隔涂层(20)以形成基因阻隔区(12)，未被阻隔涂层(20)围合的第二区域形成基因附着区(11)域；
    在所述基板(10)上加工出与所述基因附着区(11)连通的进液孔(30)和 出液孔(40)。
  12. 一种基因测序方法，其特征在于，采用权利要求10所述的基因测序芯片，所述基因测序方法包括：
    使基因测序芯片按照预定顺序依次与多种反应试剂接触，并在每次基因测序芯片与一种所述反应试剂接触后采用清洗剂对所述基因测序芯片进行清洗。
  13. 根据权利要求12所述的基因测序方法，其特征在于，使基因测序芯片按照预定顺序依次与多种反应试剂接触的步骤包括：
    针对每种反应试剂，在所述基因测序芯片与该所述反应试剂接触时，先使所述反应试剂沿所述基因测序芯片的第一导流槽(51)流动，再使所述反应试剂沿所述基因测序芯片的第二导流槽(52)流动。
  14. 根据权利要求13所述的基因测序方法，其特征在于，在所述基因测序芯片与一种所述反应试剂接触时，先使所述反应试剂沿所述基因测序芯片的第一导流槽(51)流动，再使所述反应试剂沿所述基因测序芯片的第二导流槽(52)流动的步骤包括：
    开启进液孔和第一出液孔，通过进液孔进行进液，以使反应试剂沿所述基因测序芯片的第一导流槽(51)流动；
    关闭第一出液孔，开启第二出液孔，以使反应试剂沿所述基因测序芯片的第二导流槽(52)流动。
PCT/CN2022/123237 2022-09-30 2022-09-30 基因测序芯片、载片及其加工方法以及基因测序方法 Ceased WO2024065665A1 (zh)

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