CN102703314A - Control system for DNA (Deoxyribose Nucleic Acid) sequencer - Google Patents

Abstract

The invention discloses a control system for a DNA (Deoxyribose Nucleic Acid) sequencer, which comprises a PLC (Programmable Logic Controller), controllers of first and second servo motors, first and second peristaltic pumps and driving motors of first and second multipass reversing valves, wherein the controllers of the first and second servo motors are electrically connected with the PLC; the first and second peristaltic pumps are respectively and electrically connected with the PLC and are used for receiving control signals of the PLC; a CCD (Charge Coupled Device) camera is electrically connected with the PLC and is used for receiving a control signal of the PLC; and a plurality of sensors are respectively and electrically connected with the PLC and are used for sending position signals of the CCD camera to the PLC. According to the control system disclosed by the invention, a reagent supply assembly of the DNA sequencer with a plurality of reaction bins timely and accurately supplies reagents and buffer solution for a plurality of reaction bins and the CCD camera can be ensured to timely read an optical signal in each reaction bin, and therefore, the aim of simultaneously performing a plurality of reactions is fulfilled, so that a plurality of samples can be simultaneously sequenced and the DNA sequencing efficiency is greatly improved.

Description

The system that is used for the dna sequencing appearance

Technical field

The present invention relates to the dna sequencing technical field, be specifically related to a kind of system that is used for the dna sequencing appearance.

Background technology

In the dna sequencing technical field; Tetra-sodium sequencing technologies (pyrosequencing); Be a kind of novel enzyme cascade sequencing technologies that was grown up in 1987 by people such as Nyren, its repeatable and accurate performance and Sanger method dna sequencing technology compare favourably, and speed improves greatly.

The tetra-sodium sequencing technologies is by the enzyme cascade chemiluminescence reaction in 4 kinds of enzymatic same reaction systems.The principle of tetra-sodium sequencing technologies is: after primer and the template DNA annealing; Under the synergy of archaeal dna polymerase (DNA polymerase), ATP sulfurylase (ATP sulfurylase), luciferase (luciferase) and four kinds of enzymes of apyrase (Apyrase); The polymerization of each dNTP on the primer and the release coupling of an optical signal are got up; Through detecting the release and the intensity of light, reach the purpose of The real time measure dna sequence dna.The reaction system of tetra-sodium sequencing technologies is made up of reaction substrate, strand to be measured, sequencing primer and four kinds of enzymes.Reaction substrate be 5 '-phosphinylidyne sulfuric acid (adenosine-5 '-phosphosulfate, APS) and resorcinolphthalein (luciferin).

Take turns in the sequencing reaction at each; Only add a kind of deoxynucleotide triphosphoric acid (dNTP) in the reaction system; If it can just with the next base pairing of dna profiling; Then can under the effect of archaeal dna polymerase, add 3 ' end of sequencing primer to, discharge the tetra-sodium (PPi) of a molecule simultaneously.Under the effect of ATP sulfurylase, the PPi of generation can combine to form ATP with APS, and under the catalysis of luciferase, the ATP of generation can combine to form oxyluciferin again with resorcinolphthalein, produces visible light simultaneously.Can obtain a special detected peaks through Weak light detection device and process software, the height of peak value then is directly proportional with the base number that is complementary.If the dNTP that adds can not with the next base pairing of dna profiling, then above-mentionedly instead would not take place, just do not have detected peaks yet.Remaining dNTP and residual a small amount of ATP degrade under the effect of Apyrase in the reaction system.After treating that last round of reaction is accomplished, add another kind of dNTP, above-mentioned reaction is repeated, promptly can read dna sequence dna information accurately according to the peak value figure that obtains.

The integrated operation flow process is described below: the DNA sample through fragmentation after; Application builds that storehouse reagent adds joint, microballoon, microemulsion pcr amplification, breakdown of emulsion liquid are caught, are bonded to strand; Acquisition is based upon the DNA library on the microballoon; Application add model with the laies such as enzyme of library and sequencing reaction needs to sequence testing chip with micro reaction pool, sequence testing chip and sequencing reagent are mounted on the main frame, start the order-checking program through control computer according to module number and position; Sequencing reaction is carried out in robotization; Data transmission to the data analysis computingmachine that produces is accomplished the order-checking back and is used calculating analysis software and carry out work such as picture processing, sequence are read, mass analysis, sequence assembly, finally obtains the sequence information of dna sample.The sequence testing chip that is carved with micro reaction pool carries out machine glazing core layer etching by the fibre faceplate of core diameter 25 μ m thickness 2mm and obtains, and etching depth 40 μ m amount to about 3,000,000 micro reaction pools on the chip, wherein about 1,200,000 micro reaction pools of imaging moiety.The micro reaction pool sequence testing chip is the carrier of sequencing reaction, and the DNA Beads and the various sequencing reaction that are loaded with sequencing template all are arranged in the sequence testing chip that is carved with micro reaction pool with enzyme.

In the order-checking process, chemical reaction taking place on sequence testing chip, produce visible light, catch the optical signal that sequencing reaction produced through CCD (Charge Couple Device) camera, can obtain needed order-checking information.

The contriver finds the dna sequencing appearance of prior art, has only a reaction warehouse, and an instrument can only conduct an experiment, and working efficiency is not high.In order to increase work efficiency; The contriver has made the dna sequencing appearance with a plurality of reaction warehouses, supplies reaction reagent and damping fluids for a plurality of reaction warehouses in good time and uses a CCD camera to gather the coordination problem of the optical signal in a plurality of reaction warehouses in good time because the existence of a plurality of reaction warehouses has just produced.

Summary of the invention

The technical problem that the present invention will solve is; A kind of system that is used to have the dna sequencing appearance of a plurality of reaction warehouses is provided; Control agent delivery assembly is reaction reagent and the damping fluid that each reaction warehouse provides sequencing reaction in good time, and control CCD camera motion is in time to read the optical signal in each reaction warehouse.

For addressing the above problem, the present invention adopts following technical scheme:

A kind of system that is used for the dna sequencing appearance, said dna sequencing appearance comprises a plurality of reaction warehouses; Be used to gather the CCD camera of the optical signal that dna sequencing reaction produced in each said reaction warehouse; Be used to support said CCD camera can the two dimension adjustment bracing or strutting arrangement; Said bracing or strutting arrangement comprise first servomotor that the driven CCD camera switches between different reaction warehouses with drive said CCD camera near or away from second servomotor of one of them reaction warehouse, said bracing or strutting arrangement upper edge near or be provided with a plurality of transmitters that are used to detect said CCD camera position away from the direction of said reaction warehouse; And be the agent delivery assembly that a plurality of said reaction warehouses provide reaction reagent and damping fluid; Said agent delivery assembly comprises first peristaltic pump, second peristaltic pump that is used to extract damping fluid that are used to extract sequencing reaction reagent, be used for more than first logical reversing valves of selective reagents and be used for the more than second logical reversing valves in selective reaction storehouse, and said more than first lead to reversing valves and more than second leads to reversing valves all for by motor-driven rotary valve;

Said system comprises PLC; Said PLC is electrically connected with the startup of controlling said first servomotor and second servomotor respectively, stops and turning direction with the unit of the unit of said first servomotor and second servomotor; Said PLC respectively with state first peristaltic pump, second peristaltic pump is electrically connected with the startup of controlling said first peristaltic pump and second peristaltic pump, stops and rotating speed; Said PLC is electrically connected to control the on position of logical reversing valve more than first and more than second logical reversing valves with the drive-motor of said more than first logical reversing valves and the drive-motor of more than second logical reversing valves respectively; Said PLC is electrically connected with said CCD camera to control the open and close state of said CCD camera; Said PLC also is connected with said a plurality of transmitters respectively to receive the position signal of the CCD camera that said transmitter sends.

As preferably; Said system also comprises temperature control modules; Said temperature control modules comprises the rly. that is connected with said PLC and is arranged at TP and the semiconductor heat booster in the said reaction warehouse; Said PLC receives and is connected from the signal of said TP and with the primary return of said rly., and the secondary loop of said rly. is connected in the current supply circuit of said semiconductor heat booster.

As preferably, said system also comprises the upper computer that is connected with said PLC, and said upper computer is provided with the sequence of control of said PLC operation, and transmits said sequence of control to said PLC.

As preferably, said upper computer is through RS232 port and said plc communication.

As preferably, said upper computer is communicated by letter with said CCD camera through USB port, is used to receive the view data that said CCD camera is obtained.

The system that is used for the dna sequencing appearance of the present invention; The agent delivery assembly that makes the dna sequencing appearance is that a plurality of reaction warehouses are supplied reagent and damping fluid timely and accurately; And can make the CCD camera in time read the optical signal in each reaction warehouse; Realized that a plurality of reactions carry out simultaneously, thereby can check order to a plurality of samples simultaneously, improved dna sequencing efficient greatly.

Description of drawings

Fig. 1 is the perspective view of the dna sequencing appearance of the system of employing embodiments of the invention;

Fig. 2 is the synoptic diagram of the reaction warehouse assembly of the dna sequencing appearance of the system of employing embodiments of the invention;

Fig. 3 is that the F-F of Fig. 2 is to view;

Fig. 4 is the decomposing schematic representation of one of them reaction warehouse in the reaction warehouse assembly shown in Figure 2;

Fig. 5 looks synoptic diagram for the master of the base of reaction warehouse shown in Figure 4;

Fig. 6 looks (broken section) synoptic diagram for the left side of Fig. 5;

Fig. 7 is the structural representation that the reaction tank body of reaction warehouse shown in Figure 4 cooperates with tail rod;

Fig. 8 looks (broken section) synoptic diagram for the left side of Fig. 7;

Fig. 9 is the structural representation of the reaction tank body among Fig. 8;

Figure 10 is the structural representation of the tail rod among Fig. 8;

Figure 11 is that the A-A of Fig. 7 is to sectional drawing;

Figure 12 looks synoptic diagram for the master of the mount pad of reaction warehouse shown in Figure 4;

Figure 13 looks (broken section) synoptic diagram for the left side of Figure 12;

Figure 14 is that the B-B of Figure 12 is to sectional schematic diagram;

Figure 15 is that the C-C of Figure 14 is to synoptic diagram;

Figure 16 is the front view of the slip hyoplastron among Figure 14;

Figure 17 is the left view of Figure 16;

Figure 18 is the perspective view of the baffle plate among Figure 14;

Figure 19 is the side-view of baffle plate shown in Figure 180;

Figure 20 is that the D-D of Figure 19 is to sectional view;

Figure 21 is the structural representation (front end of camera body is connected with joint flange) of the CCD camera of the dna sequencing appearance of the system of employing embodiments of the invention;

Figure 22 is the front view of the joint flange among Figure 21;

Figure 23 is that the E-E of Figure 22 is to sectional view;

Structural representation (broken section) when Figure 24 is connected with the CCD camera for one of them reaction warehouse of the dna sequencing appearance of the system that adopts embodiments of the invention;

Figure 25 for the dna sequencing appearance of the system that adopts embodiments of the invention can the two dimension adjustment the perspective view that is connected with the CCD camera of bracing or strutting arrangement;

Figure 26 be among Figure 25 can two dimension adjustment the perspective view of second straight-line motion mechanism of bracing or strutting arrangement;

Figure 27 is the vertical view of second straight-line motion mechanism shown in Figure 26;

Figure 28 be Figure 27 along G-G to sectional view;

Figure 29 be Figure 28 along H-H to sectional view;

Figure 30 is the liquid line structure synoptic diagram of agent delivery assembly of dna sequencing appearance of the system of embodiments of the invention:

Figure 31 is an agent delivery assembly control method schema shown in Figure 30:

Figure 32 is the structural representation of the system that is used for the dna sequencing appearance of embodiments of the invention:

Figure 33 is the structural representation (is example with a reaction warehouse) of the temperature control modules of the system that is used for the dna sequencing appearance of embodiments of the invention.

Main description of reference numerals

101-brace table 103-vibroshock 102-vibration damper plate

104-reaction warehouse assembly 110-CCD camera 105-can the two-dimentional bracing or strutting arrangement of adjusting

1-camera body 2-joint flange 4-fibre faceplate

5-reaction tank body 6-mount pad 7-sequence testing chip

The protruding circle of 8-base 9-tail rod 21-

41-screw 44-through hole 45-cover

49-the 3rd step 51-sequencing reaction pond 54-backboard

55-boss 56-annular recesses 57-heating chamber

58-sealing-ring 61-screw 62-slip hyoplastron

63-pedestal 64-baffle plate 66-ring groove

67-retaining ring 69-installation cavity 80-cavity

The 81-first spring 82-second through hole 84-bracing frame

85-spring lever 86-spring support 87-rotating shaft

91-connection section 92-sliding part 93-boss

The 31-first straight-line motion mechanism 32-second straight-line motion mechanism

35-supporting seat 38-transmitter

The 271-second leading screw 272-nut 281-limit base

The 311-first track base 312-first guide rail 313-first slide block

The 319-first limited block 321-second track base 322-second guide rail

The 323-second slide block 326-second servomotor 327-shaft coupling

328-gag lever post 380-groove 381-shading piece

510-fluid inlet 511-liquid outlet 561-TP

571-semiconductor heat booster 572-shut 605-glissile segment

606-construction section 610-first step 612-screw

621-tuck 623-second step 633-second spring

The 700-main pipe line 701-first peristaltic pump 702-more than first logical reversing valves

The 711-second peristaltic pump 712-more than the second logical reversing valve 731-first froth in vacuum devices

The 732-second froth in vacuum device 741-reagent bottle 742-waste liquid barrel

743 damping fluid bottles, 750 reaction warehouse 760-three-way connectors

The 841-screw

Embodiment

Below in conjunction with accompanying drawing and specific embodiment the present invention is described in further detail, but not as to qualification of the present invention.

At first combine accompanying drawing 1-Figure 29 to specify the detailed structure of the dna sequencing appearance of the system that adopts embodiments of the invention.The system of present embodiment can be applied in the dna sequencing appearance with a plurality of reaction warehouses, is not limited to the described dna sequencing appearance of present embodiment.

Like Fig. 1, Fig. 2 and shown in Figure 3, the applied dna sequencing appearance of system of the present invention comprises brace table 101, a plurality of vibroshock 103 and the vibration damper plate 102 that is connected with brace table 101 through a plurality of vibroshocks 103; Be used to carry out the reaction warehouse assembly 104 of dna sequencing reaction, reaction warehouse assembly 104 comprises the bracing frame 84 that vertically is arranged on the vibration damper plate and is set up in parallel a plurality of reaction warehouses on bracing frame 84; In the present embodiment, be that example describes with four reaction warehouses.Be used to gather the CCD camera 110 of the optical signal that dna sequencing reaction produced in the reaction warehouse; Be used to support CCD camera 110 and drive CCD camera 110 when aiming at one of them reaction warehouse near or away from every reaction warehouse, and and the position that aligns of each reaction warehouse between switch can the two dimension adjustment bracing or strutting arrangement 105; Being used to reaction warehouse assembly 104 provides the system 109 of reaction reagent and damping fluid; Reaction warehouse assembly 104 all is fixedly installed on the vibration damper plate 102 with bracing or strutting arrangement 105 that can the two dimension adjustment, and system 109 is arranged on the brace table 101.In the present embodiment; Brace table 101 is positioned on the cabinet (not shown), and instrument and circuit element that sequencing analysis is used are set in the cabinet, and vibroshock 103 adopts the air damp type vibroshock; Reached the isolation external shock; Eliminate or weaken its influence, in the present embodiment, respectively settled a vibroshock 103 for four of 102 lower ends jiaos at the bottom of the damping dna sequencing appearance workmanship.

At first combine Fig. 1-Figure 24 that the detailed structure and the working process thereof of reaction warehouse assembly 104 of the applied dna sequencing appearance of system of present embodiment is described.

Like Fig. 2, Fig. 3, Fig. 4 and shown in Figure 24, in the present embodiment, each reaction warehouse includes base 8, the reaction tank body 5 that carries out sequencing reaction that is rotatably connected on the bracing frame 84 and is used to install the mount pad 6 of the sequence testing chip of sequence testing chip 7; The mount pad 6 of sequence testing chip is fixedly connected on the reaction tank body 5; Base 8 and reaction tank body 5 are provided with and pass its two a plurality of tail rods 9; And tail rod 9 is fixedly connected with base 8; Reaction tank body 5 is enclosed within on the tail rod 9, tail rod 9 axially on slide and the fine motion that can make progress in the footpath of tail rod 9.Be provided with first elastomeric element between reaction tank body 5 and the base 8, as shown in Figure 1, in the present embodiment; First elastomeric element is preferably first spring 81; Spring has simple in structure, and the advantage of being convenient to install can also select for use other elastomeric elements that can realize said function to realize certainly.

Like Fig. 4, Fig. 5 and shown in Figure 6, a side of base 8 is provided with cavity 80, is provided with two spring levers 85 in the cavity 80 side by side, and spring lever 85 integral body are positioned at cavity 80, can not stretch out cavity 80.Each spring lever 85 is attend sheathed one first spring, 81, the first springs 81 of branch and is stretched out cavity 80.As preferably, in the present embodiment, the bottom of cavity 80 is provided with spring support 86, and spring lever 85 is positioned on the spring support 86, and the spring support 86 of increase can reduce the required length of first spring 81.It is for the ease of miscellaneous part is installed that cavity 80 is set.

Like Fig. 4, Fig. 7 and shown in Figure 11; Reaction tank body 5 comprises backboard 54 and is located on the said backboard 54 and the boss 55 of the side protrusion of toward back plate 54; The end face of said boss 55 is provided with the annular recesses 56 of sealing, and the zone that the annular recesses 56 of sealing is surrounded is sequencing reaction pond 51; As shown in Figure 7, be embedded with sealing-ring 58 in the annular recesses 56, the end face of sealing-ring 58 protrusion boss 55, to surround sequencing reaction pond 51, wherein the end face of boss 55 is smooth as much as possible.Like Fig. 4, Fig. 7, Fig. 8 and shown in Figure 9; The side that reaction tank body 5 is provided with backboard 54 is attached on the base 8, and the boss 55 of reaction tank body 5 is provided with four first through holes, and said first through hole is step-like; One section near base is glissile segment 605; Another section of first through hole is a construction section 606, and the internal diameter of construction section 606 forms first step 610 greater than glissile segment 605.Like Fig. 7-shown in Figure 10; One end of tail rod 9 is the connection section 91 that is connected with base 8; The other end of tail rod 9 is provided with the boss 93 that is used to be held in first step 610 places, and the middle part of tail rod 9 is to be used for the sliding part 92 that cooperates with glissile segment 605 near the part of boss 93.Shown in figure 12, connection section 91 is provided with screw thread, and is as shown in Figure 7, and the end of the boss 93 of tail rod 9 is provided with the cutter groove that is used to cooperate screwdriver.As shown in Figure 1, tail rod 9 passes first through hole and second through hole 82 on the base 8 on the reaction tank body 5, and the back that makes connection section 91 expose base 8 is fastening through nut then, thereby realizes being fixedly connected between tail rod 9 and the base 8.Realize being slidingly connected in order to make between reaction tank body 5 and the tail rod 9; The somewhat larger in diameter of glissile segment 605 is in the diameter of sliding part 92; Like this reaction tank body 5 both can be implemented in tail rod 9 axially on slip, the fine motion that again can realization response pond body 5 makes progress in the footpath of tail rod 9.In the present embodiment, as preferred version, tail rod 9 is provided with boss 93; So that reaction tank body 5 is being slidingly connected at 9 last times of tail rod; Through boss 93 position-limiting action with first step 610, tighten with connection section 91 on during the nut of thread fit, boss 93 driving a reaction pond bodies 5 move to the direction of base 8; To compress first spring 81; Make between reaction tank body 5 and the base 8 to have certain preload, it is exactly in order to be attached to 8 last times of base when reaction tank body 5 that above-mentioned first spring 81 stretches out cavity 80, and first spring 81 can be given reaction tank body 5 certain function power.

Like Fig. 4, Figure 12, Figure 13 and shown in Figure 14; The mount pad 6 of sequence testing chip comprises pedestal 63 and is opened in the installation cavity 69 on the pedestal 63; The mount pad 6 of sequence testing chip is positioned at the side that reaction tank body 5 is provided with boss 55; Being fixedly connected between pedestal 63 and the backboard 54 stretches in the installation cavity 69 boss 55; Sequence testing chip 7 is fixed in the outer end of installation cavity 69, is provided with the gap that supplies order-checking to flow through with reagent between the end face of sequence testing chip 7 and boss 55, and this gap is through being embedded at sealing-ring 58 sealings in the annular recesses 56; The size in said gap is about 0.2mm, and sealing-ring 58 convex hole connected in stars 56 contact the sequencing reaction pond 51 with the confession sequencing reaction that surrounds sealing with sequence testing chip 7.

Like Fig. 7, shown in Figure 11; In the present embodiment, this annular recesses 56 is a hexagon, and hexagonal two ends have fluid inlet 510 and liquid outlet 511; When reaction tank body 5 is vertically placed; Fluid inlet 510 is in hexagonal lower end, and liquid outlet 511 is in hexagonal upper end, forms the flow hydrokinetics advantageous shape that balanced flow is crossed in reaction warehouse.Shown in figure 11; Bottom in sequencing reaction pond 51 is fixed with TP 561 through heat conductive silica gel, is not provided with the opposite side of annular recesses 56 at reaction tank body 5, offers the heating chamber 57 that is used for ccontaining semiconductor heat booster 571; Heating chamber 57 is opened on the side of reaction tank body 5; Fixing semiconductor heat booster 571 on its bottom surface with to 5 heating of reaction tank body, guarantees that the temperature that is positioned at annular recesses 56 is controlled at 35 ± 1 ℃.In the present embodiment, heating chamber 57 is through shut 572 sealings, so that be adjacent on the mount pad 6 of sequence testing chip with reaction tank body 5.

Like Fig. 4, Figure 12 and shown in Figure 13; The pedestal 63 of the mount pad 6 of sequence testing chip is provided with four third through-holes; Four set screws 612 are connected on the backboard 54 of four screws 61 of the mount pad 6 of sequence testing chip through passing these four third through-holes and reaction tank body 5, thereby realize being fixedly connected between the backboard 54 of pedestal 63 and reaction tank body 5 of mount pad 6 of sequence testing chip.Like Figure 12-shown in Figure 15; The pedestal 63 of the mount pad 6 of sequence testing chip is provided with and comprises the installation cavity 69 that is used for ccontaining sequence testing chip 7; One end of installation cavity 69 is provided with the retaining ring 67 that is used to locate sequence testing chip 7; It radially is provided with locating slot the inwall upper edge of installation cavity 69, be provided with in the locating slot along locating slot slide stretch in the installation cavity 69 with hold sequence testing chip 7 make it be attached on the retaining ring 67 or the withdrawal locating slot in to unclamp the slip hyoplastron 62 of sequence testing chip 7.When slip hyoplastron 62 stretches out and the distance between the retaining ring 67 equal the thickness of sequence testing chip just, sequence testing chip 7 is adjacent on retaining ring 67.The locating slot symmetry is located on the inwall of installation cavity 69 and connects its inwall, and its quantity can be even numbers such as two or four, in order stable holding to live sequence testing chip 7, and the circumferential direction uniform distribution of a plurality of locating slots on the axial same position of installation cavity 69.Certainly, more a plurality of locating slots can also be set, but more locating slot can cause the inconvenience of operation in order to improve holding force.In the present embodiment, locating slot is two, and for easy to process, locating slot is a circle hole shape.As shown in Figure 5, locating slot is less near one section size of installation cavity 69, is formed for the second step 623 of spacing slip hyoplastron 62; Be locating slot near a section of installation cavity 69 be a less circular hole, rest part is a bigger circular hole.Like Fig. 9 and shown in Figure 10, slip hyoplastron 62 is provided with the tuck 621 of protrusion.As shown in Figure 7, when slip hyoplastron 62 when installation cavity 69 stretches into, the tuck 621 of slip hyoplastron 62 is resisted against said second step 623 places, an end of slip hyoplastron 62 stretches in the installation cavity 69 to hold sequence testing chip 7.The outer end of locating slot is provided with the baffle plate 64 that is fixedly connected with mount pad 6; Between the tuck 621 of slip hyoplastron 62 and baffle plate 64, be provided with second spring 633; In the present embodiment; Second spring 633 is enclosed within the other end of slip hyoplastron 62, and the two ends of second spring 633 are resisted against respectively on tuck 621 and the baffle plate 64.Like Figure 13, shown in Figure 14, on the surface of the mount pad 6 of sequence testing chip, have ring groove 66, ring groove 66 is centered around the outside of installation cavity 69, makes sequence testing chip 7 be positioned at the zone that ring groove 66 is surrounded.

As shown in Figure 8, baffle plate 64 is fixedly connected with mount pad 6 through two screws 41, is convenient to installation and removal.Second spring 633 is enclosed within on the slip hyoplastron 62 between tuck 621 and the baffle plate 64; Like Figure 11-shown in Figure 13; Baffle plate 64 is provided with the through hole 44 that supplies slip hyoplastron 62 to pass away from an end of installation cavity 69; The inboard of baffle plate 64 is provided with the cover 45 of internal diameter greater than through hole 44, the end that is formed for making second spring 633 between cover 45 and the through hole 44 against the 3rd step 49.In the present embodiment, second spring 633 can also adopt other second elastomeric elements to replace, and baffle plate 64 can also adopt miscellaneous part, as long as can be against the other end of second spring 633.When sequence testing chip 7 is installed, stir the end in the installation cavity 69 that extend into of slip hyoplastron 62, make in its withdrawal locating slot; Tuck 621 extruding second spring 633 of slip hyoplastron 62; Unclamp slip hyoplastron 62 after putting into sequence testing chip 7 installation cavity 69 and be attached on the retaining ring 67, slip hyoplastron 62 stretches out under the effect of second spring 633, is clamped in the side of sequence testing chip 7; Sequence testing chip 7 is adjacent on retaining ring 67, even also unlikelyly when guaranteeing that sequence testing chip 7 is inverted deviate from.

For the ease of loading and unloading sequence testing chip 7,, as shown in Figure 5 as preferably; Dna sequencing appearance in the present embodiment comprises bracing frame 84, and base 8 is through being rotatably connected on the bracing frame 84 like Fig. 4 and rotating shaft 87 shown in Figure 6, when installing or unloading sequence testing chip 7; Let base 8 be positioned at level attitude, when carrying out sequencing reaction, base 8 is positioned at vertical position as shown in Figure 5; Base 8 is fixedly connected with bracing frame 84 through screw 841, an end band knob of screw 841, and the other end is provided with connecting thread.

Shown in figure 21; Comprise camera body 1 and the fibre faceplate 4 that is used to read the optical signal that occurs in the sequencing reaction on the said sequence testing chip 7 that is positioned at said camera body 1 with the CCD camera of the dna sequencing appearance of present embodiment coupling, the side that an end of fibre faceplate 4 stretches out the sequence testing chip 7 of the direct mount pad 6 (structure of the mount pad 6 of sequence testing chip sees above and is described in detail) with the sequence testing chip that is positioned at the dna sequencing appearance of the front end of said camera body 1 directly contacts to obtain the optical signal that sequencing reaction was produced on the opposite side that occurs in sequence testing chip 7.In the present embodiment, as preferably, the optical fiber core diameter of said fibre faceplate is 6 μ m, and the optical fiber core diameter can be selected in 1-15 μ m.Fibre faceplate 4 directly stretches out the front end of camera body 1, does not use camera lens, and fibre faceplate 4 can directly be coupled with sequence testing chip 7, simplifies light path, reduces light loss, improves the order-checking quality of signals.

Like Figure 21, shown in Figure 24, in the present embodiment, as preferably, the front end of camera body 1 is connected with joint flange 2.Shown in Figure 21, Figure 22 Figure 52 3, the front end of camera body 1 have the ring groove 66 that is used to insert on the mount pad 6 of being located at sequence testing chip and with ring groove 66 close-fitting protruding circles 21.Joint flange 2 and camera body 1 are connected through bolt, and the fibre faceplate 4 in the camera body 1 is positioned at the zone that protruding circle 21 is surrounded, and promptly protruding circle 21 stretches out the outside that joint flange 2 is centered around fibre faceplate 4.The end face of fibre faceplate 4 stretches out protruding circle 21 certain distances; Can between 1-3mm, adjust; In the present embodiment; The distance that the end face of fibre faceplate 4 stretches out said protruding circle 21 is 2mm, directly contacts with sequence testing chip 7 so that the end face of fibre faceplate 4 can stretch in the installation cavity 69 of mount pad 6 of sequence testing chip.Present embodiment through be provided with can with the direct-coupled fibre faceplate 4 of sequence testing chip; The faint visible light that sequencing reaction produced that on sequence testing chip 7, carries out is directly received by fibre faceplate 4; Be converted into electrical signal; Improve coupling efficiency, coupling efficiency can surpass 70%, has guaranteed obtaining of high-quality order-checking signal.In addition, ring groove 66 is complementary with the shape of protruding circle 21, makes protruding circle 21 insert can form in the ring groove 66 and is sealed and matched.Shown in figure 24; When the CCD camera during near the mount pad 6 of sequence testing chip; The joint flange 2 that is installed in camera body 1 front end is near the sequence testing chip 7 on the mount pad 6 of sequence testing chip, and in the ring groove 66 on the mount pad 6 of the protruding circle 21 entering sequence testing chips of the ring-type on the joint flange 2, protruding circle 21 is complementary with the shape of ring groove 66; The formation sealed structure can closely cooperate; Thereby form the darkroom environment of light stopping property, the luminous energy that further guarantees sequencing reaction and produced is received by fibre faceplate 4, guarantees obtaining of order-checking signal.Through have the joint flange 2 of protruding circle 21 in camera body 1 front end increase; And on the mount pad 6 of sequence testing chip, offer corresponding annular groove 66; Thereby between the mount pad 6 of sequence testing chip and camera body 1, form the darkroom environment, when guaranteeing fibre faceplate 4 couplings of sequence testing chip 7 and the front end that is positioned at camera body 1, not disturbed by extraneous light; Thereby reduced ground unrest, further guaranteed the obtaining of order-checking signal of high s/n ratio.

Structure when the reaction warehouse that Figure 24 shows the applied dna sequencing appearance of system of present embodiment is connected with the CCD camera, the structural representation when promptly the dna sequencing appearance is in running order.Shown in figure 24; When carrying out sequencing reaction; The camera body 1 of CCD camera is near reaction warehouse, and the outer face of exposing the fibre faceplate 4 of camera body 1 front end is attached on the sequence testing chip 7, along with the direction of camera body 1 to reaction warehouse moves; Fibre faceplate 4 promotes sequence testing chip 7 and moves; Because sequence testing chip 7 is fixed in the mount pad 6 of sequence testing chip, also be fixed connection between the mount pad 6 of sequence testing chip and the reaction tank body 5, thereby sequence testing chip 7 can drive reaction tank body 5 endwisely slipping at tail rod 9 upper edge tail rods 9; Because reaction tank body 5 receives the reactive force of first spring 81 between reaction tank body 5 and the base 8, is close to each other between the outer face of sequence testing chip 7 and the fibre faceplate 4.In the outer face of sequence testing chip 7 with fibre faceplate 4 do not realize fully parallel before; Owing to receive the reactive force of first spring 81; Be close in the process of sequence testing chip 7 in the outer face of fibre faceplate 4, reaction tank body 5 is when receiving the reactive force of first spring 81, and fine motion makes progress in the footpath of tail rod 9; Drive sequence testing chip 7 fine motion that makes progress in the footpath of tail rod 9, thereby realize the outer face of fibre faceplate 4 and the adjustment of the parallelism between the sequence testing chip 7.Sequence testing chip 7 is through in the adjustment of tail rod 9 on axially; Realized pressing close to as much as possible between the outer face of outer face and fibre faceplate 4 of sequence testing chip 7; Reaction tank body 5 fine motion that makes progress in the footpath of tail rod 9; Realized having the high parallelism precision of trying one's best between outer face and the sequence testing chip 7 of fibre faceplate 4, guaranteed to greatest extent at fibre faceplate 4 to have prevented external optical pollution when sequence testing chip 7 reads the fluorescent signal of sequencing reaction; Thereby reduced ground unrest, realized obtaining of high s/n ratio order-checking signal.

Explain below in conjunction with Fig. 1, Figure 25, Figure 26 present embodiment the applied dna sequencing appearance of system can two dimension adjustment the structure and the working process thereof of bracing or strutting arrangement.

Like Fig. 1, Figure 25, shown in Figure 26; The applied dna sequencing appearance of the system of present embodiment can two dimension adjustment bracing or strutting arrangement; Comprise be used to drive first straight-line motion mechanism 31 that CCD camera 110 switches between the mount pad of a plurality of reaction warehouses arranged side by side of reaction warehouse assembly 104 with drive CCD camera 110 near or away from second straight-line motion mechanism 32 of each reaction warehouse; Wherein, First straight-line motion mechanism 31 comprises first track base 311, be located at first guide rail 312 on first track base 311, along first guide rail, 312 straight-line first slide blocks 313 be positioned at the first ball-screw kinematic pair of first track base 311, the nut of the first ball-screw kinematic pair (not shown among Figure 25) is fixedly connected with first slide block 313; Second straight-line motion mechanism 32 comprises second track base 321, be located at second guide rail 322 on second track base 321, along second guide rail, 322 straight-line second slide blocks 323 be positioned at the second ball-screw kinematic pair of second track base 321; The nut of the second ball-screw kinematic pair (not shown among Figure 25, the description that sees below) is fixedly connected with second slide block 323; Second track base 321 is fixedly connected with first slide block 313 and second guide rail 322 and the 312 vertical settings of first guide rail, and second slide block 323 is fixedly connected with the supporting base 35 of the CCD camera 110 that is positioned at its top.Because second guide rail 322 and the 312 vertical settings of first guide rail; Second straight-line motion mechanism 32 and the CCD camera 110 that is fixedly connected with it are when first guide rail 312 moves; Just can between the differential responses storehouse, switch; When second straight-line motion mechanism 32 moves to the mount pad over against some reaction warehouses, under the drive of second straight-line motion mechanism 32, CCD camera 110 just towards or away from the direction motion of the mount pad of this reaction warehouse.The mount pad in CCD camera 110 orientating reaction storehouses motion and when being adjacent to mount pad; Carry out examining order; Optical signal in the catching reaction storehouse; CCD camera 110 just can switch between the mount pad of different reaction warehouses under the drive of first straight-line motion mechanism 31 during away from the mount pad of reaction warehouse.

First straight-line motion mechanism 31 and second straight-line motion mechanism, 32 structures are basic identical, and different is, what the upper fixed of first slide block 313 connected is second track base 321, are the supporting bases 35 of CCD camera 110 and the upper fixed of second slide block 23 connects.Specify the structure and the working process of second straight-line motion mechanism 32 below in conjunction with Figure 26-Figure 29, need to prove,, equally also can be applied on first straight-line motion mechanism 31 any preferred implementation of second straight-line motion mechanism 32.

Like Figure 26-shown in Figure 29; Second straight-line motion mechanism 32 also comprises second servomotor 326 that is connected with second leading screw 271 of the second ball-screw kinematic pair; Certainly, first straight-line motion mechanism also comprises the first servomotor (not shown) that is connected with first leading screw of the first ball-screw kinematic pair.Second straight-line motion mechanism 32 is realized straight-line through two second guide rails 322 that are set in parallel on second track base 321; Second slide block 323 slides on second guide rail 322 and has guaranteed its straight-line precision; Second servomotor 326 drives the slip of second slide block 323 on second guide rail 322 through the second ball-screw kinematic pair; Wherein, Second leading screw 271 of the second ball-screw kinematic pair is between two second guide rails 322, and the nut 272 of the second ball-screw kinematic pair is fixedly connected with second slide block 323, can guarantee the precision of second straight-line motion mechanism 32 like this.Like Figure 27, shown in Figure 28, be connected through shaft coupling 273 between second leading screw 271 of the second ball-screw kinematic pair and second servomotor 326.

Shown in Figure 26-29, a side of second track base 321 is provided with a plurality of transmitters 38 along the length direction of second guide rail 322.Transmitter 38 is a photo-sensor, and a side of second track base 321 is provided with five grooves 380 side by side along the length direction of second guide rail 322, and the opening of groove 380 upwards; The length direction of groove 380 is along the length direction setting of second guide rail 322; One side of groove 380 is an emission side, and opposite side is fixedly connected with the shading piece 381 that can from groove 380, pass through for the induction side on second slide block 323; Like Figure 26, shown in Figure 29, shading piece 381 is one " L " shape board.When 323 motions of second slide block, when drive shading piece 381 passed through each groove 380 successively, the induction side of corresponding grooves 380 was just sent corresponding signal to gear.The position of groove 380 on second guide rail, 322 length directions is according to the actual needs setting; Be provided with five grooves in the present embodiment; The groove at two ends is corresponding respectively allow to leave or near mount pad farthest and closest range; When shading piece 381 reached these two positions, gear just sent guard signal.Three positions of the corresponding second servomotor works better of three grooves of intermediary, when shading piece 381 reached these three positions, induction side direction gear sent corresponding signal.Certainly, a side of first track base 311 transmitter that can respective numbers also be set according to the quantity and the spacing needs of mount pad.

Like Fig. 1, shown in Figure 25; The end that one end of first track base 311 is provided with first limited block, 319, the second track bases 321 is provided with second limited block 329, and the other end is provided with gag lever post 328; Second track base 321 is fixed with limit base 281 near an end of mount pad; Gag lever post 328 is located in the limit base 281 along the length direction of second guide rail 322, and between end of gag lever post 328 and limit base 281, has been provided with the spring of shock absorption, when transmitter breaks down; When second slide block 323 can not stop to the direction motion of bracing frame 84 automatically; Stop it to move on by gag lever post 328 and limit base 281, avoid CCD camera 100 and 104 collisions of reaction warehouse assembly, prevent to damage.

Specify the working process of the agent delivery assembly of present embodiment below in conjunction with Fig. 1, Figure 30 and Figure 31.

Shown in figure 30, the agent delivery assembly comprises: be used for the buffering pipe 720 of the preparation pipeline 710 of a complete set of reaction reagent of the ccontaining question response of arranging in order, the damping fluid that is used to flow through and be used to the main pipe line 700 of the mixed solution of said reaction warehouse supply reaction reagent and damping fluid; Said preparation pipeline 710, buffering pipe 720 are connected through a three-way connector 760 with main pipe line 700.A complete set of reaction reagent refers to all reagent that carry out the single reaction in the reaction warehouse.

Be used to extract first peristaltic pump 701 of sequencing reaction reagent; Have more than first logical reversing valves 702 of a plurality of imports and an outlet, have more than second of an import and a plurality of outlets and lead to reversing valves 712; Each import of logical reversing valve 702 more than first is communicated with a reagent bottle 741 respectively, and the outlet of logical reversing valve 702 more than first is connected with preparation pipeline 710 through first peristaltic pump 701; Be used for being connected with buffering pipe 720 from second peristaltic pump, 711, the second peristaltic pumps 711 that damping fluid bottle 743 extracts damping fluid.The import of logical reversing valve 712 more than second is communicated with main pipe line 700, and an outlet of logical reversing valve 712 more than second is communicated with waste liquid barrel 742, is used for bypass; The outlet of other of logical reversing valve 712 more than second is communicated with the fluid inlet of a reaction warehouse 750 respectively; The liquid outlet of each reaction warehouse 750 is communicated with waste liquid barrel.Shown in figure 30; Whole liquid road relies on first peristaltic pump 701 and second peristaltic pump 711 that flow stream pressure is provided; First peristaltic pump 701 and second peristaltic pump 711 are respectively by corresponding step motor drive; Different reagent are selected through more than first logical reversing valves 702 according to the sequential of programdesign successively; Through three-way connector 760 time, converge dilution with damping fluid, get into main pipe line 700, get into one of four reaction warehouses 750 or bypass liquid road (bypass liquid road is exactly directly to get into waste liquid barrel 742 from more than second logical reversing valves 712 without reaction warehouse 750) respectively through more than second logical reversing valves 712.As preferably, in the present embodiment, main pipe line 700 is provided with the first froth in vacuum device 731, and to subdue dissolved gases, accurately control gets into reagent and the amount of damping fluid in the reaction warehouse.In order to eliminate gas dissolved and bubble in the liquid road better, because the flow velocity of damping fluid is very fast, reach 4mL/min, more be prone to produce bubble, on the pipeline between the damping fluid bottle 743 and second peristaltic pump 711, be provided with the second froth in vacuum device 732.The second froth in vacuum device 732 is removed bubble and the most of gas dissolved that exists in the damping fluid.The first froth in vacuum device 731 and the second froth in vacuum device 732 all provide the gas negative pressure through connected vacuum pump.Logical reversing valve 702 more than first and more than second logical reversing valves 712 can adopt SV.

Reagent bottle 741 has ten; Wherein nine kinds of reagent such as nine dATP that are used to hold the dCTP that carries out sequencing reaction, dGTP, dTTP, α position sulfo-, ATP, substrate (resorcinolphthalein and APS), apyrase, apyrase suppressor factor also have a reagent bottle to be used to hold the damping fluid that is used to isolate said reagent.(damping fluid in this reagent bottle is used to isolate adjacent reagent, and is different with the effect of damping fluid in the damping fluid bottle 743), holds dilution in the damping fluid bottle 743 and clean the damping fluid of usefulness.Wherein, Reagent such as the dATP of dCTP, dGTP, dTTP, α position sulfo-, ATP, substrate (resorcinolphthalein and APS), apyrase and apyrase suppressor factor are connected with the import of more than first logical reversing valves 702 respectively; In the present embodiment; Logical reversing valve 702 more than first and more than second logical reversing valves 712 are ten 11 logical valves, so that reserve subsequent use passage.According to test reaction sequential, the outlet of logical reversing valve 702 more than first is communicated with one of them reagent bottle, drive by first peristaltic pump 701, and flow velocity 0.7mL/min, front and back first peristaltic pump 701 that at every turn switches reagent all stops 1s, to avoid the crossed contamination of reagent.

Damping fluid in the damping fluid bottle 743 is driven by second peristaltic valve 731, gets into three-way connector 760, goes into main pipe line 700 with the reagent mix dilution is laggard.The pipeline that is exported to three-way connector 760 inlets of first peristaltic pump 701; The length of promptly preparing pipeline 710 is 285mm; Carrying out a complete set of reagent of single reaction in reaction warehouse will arrange in preparation pipeline 710 according to the order of sequence; Get into three-way connector 760 then successively; With inject reaction warehouse 750 continuously through main pipe line 700 and follow-up liquid road after damping fluid mixes and participate in sequencing reaction, avoided in primary first-order equation, need repeatedly stopping the situation of second peristaltic pump 711, solved because the reagent of secondary response lacks power and causes the uneven problem of reacting before in the termination of pumping process in reaction warehouse.In the present embodiment, the length of preparation pipeline 710 can be carried out a complete set of reagent that single reacts in the ccontaining reaction warehouse at least.Mixed solution in the main pipe line 700 gets into more than second logical reversing valves 712, and through the fluid inlet entering reaction warehouse 750 of program selection from one of them reaction warehouse 750, carries out sequencing reaction, and the liquid outlet of reaction warehouse 750 is communicated with waste liquid barrel 742.If what carry out is matting, the liquid that then comes out from more than second logical reversing valves 712 directly gets into bypass liquid road, and waste liquid barrel 742 is directly connected on bypass liquid road; In the present embodiment; Four outlets of logical reversing valve 712 more than second are communicated with the fluid inlet of four reaction warehouses 750 respectively, and an outlet of logical reversing valve 712 more than second is communicated with waste liquid barrel 742 through bypass liquid road, so that use the damping fluid flushing pipe; Other four outlets are reserved with reaction warehouse 750 and are communicated with, and flow out for reaction reagent.

In order to realize effect preferably, the length of preparation pipeline 710 should be sufficiently long to a complete set of reagent (nine kinds) that can hold the single reaction.As preferred version, the length of preparation pipeline 710 also should be sufficiently long to the damping fluid between the different reagent in a complete set of reagent that holds single reaction and a complete set of reagent.

Below in conjunction with schema shown in Figure 31, specify the control method of above-mentioned agent delivery assembly, comprise the steps:

S1: open said first peristaltic pump;

S2: according to the order of sequencing reaction; Open an import of said more than first logical reversing valves; The said reagent bottle of its outlet first kind reagent required with being contained with sequencing reaction is communicated with, and the reagent that extracts in the said reagent bottle gets into the preparation pipeline that is connected with the said first wriggling pump outlet, and the extraction order of reagent is carried out according to the TR of sequencing reaction; The reaction reagent that each reagent bottle is corresponding different, and corresponding with an import of said more than first logical reversing valves;

S3: when first kind of reagent reaches predetermined amount, stopped for first peristaltic pump for some time,, can select between second, avoid the interference between the adjacent reagent at 1-2 like 1s;

S4: open said first peristaltic pump; Open other imports of said more than first logical reversing valves successively and extract other reagent respectively and get into said preparation pipeline; And when every kind of reagent reaches predetermined amount; Stop said first peristaltic pump for some time, in a reaction warehouse, carry out a complete set of reagent of single reaction and all send into till the said preparation pipeline;

Order according to sequencing reaction; Extract needed second kind of reagent; And then stop first peristaltic pump, extract then the third reagent, the 4th kind of reagent ..., so circulation; In a needed reaction warehouse, carry out a complete set of reagent of single reaction and all send into said preparation pipeline; In the present embodiment, totally nine kinds of a complete set of reagent are dATP, ATP, resorcinolphthalein, APS, apyrase and the apyrase suppressor factor of nine kinds of reagent: dCTP carrying out sequencing reaction, dGTP, dTTP, α position sulfo-successively.The amount of every kind of reagent; Promptly the opening time of first peristaltic pump is provided with according to the requirement of sequencing reaction in advance; It is for realize can be in a reaction warehouse carries out the cycle of once sequencing reaction that a complete set of reagent all is arranged on the said preparation pipeline; Needn't stop any pump, avoid the liquid in the place ahead that termination of pumping causes to lack power, reduce flow velocity and influence the sequencing reaction quality in the reaction warehouse; In order to prevent adjacent two kinds of reagent phases mutual interference; In the present embodiment; After accomplishing any one reagent of extraction, open said first peristaltic pump, the import of said more than first logical reversing valves is communicated with a reagent bottle that holds damping fluid (wherein having a reagent bottle to be contained with damping fluid); Be used to extract damping fluid and form fluid column, this fluid column between two kinds of adjacent reagent to isolate two kinds of adjacent reagent.

S5: open said first peristaltic pump and said second peristaltic pump that is used to extract damping fluid simultaneously, said a complete set of reagent gets into main pipe line with the three-way connector that damping fluid process together and said first peristaltic pump are connected with second peristaltic pump;

S6 a: outlet of opening said more than second logical reversing valves; Be positioned at the reagent of main pipe line and the mixed solution of damping fluid and get into corresponding reaction warehouse; The opening sequence of logical reversing valve is provided with according to the reaction sequence of reaction warehouse more than second; In the present embodiment, four reaction warehouses are first, second, third, fourth reaction warehouse according to its position number consecutively.

Just realized being followed successively by the purpose that each reaction warehouse provides a complete set of reagent according to above-mentioned control method.

As priority scheme, the control method of the agent delivery assembly of present embodiment also comprises the steps:

S7: judge whether a complete set of reagent that a reaction warehouse carries out single reaction all gets into this reaction warehouse, if, performing step S8;

S8: the outlet that said more than second logical reversing valves are communicated with waste liquid barrel is communicated with the import of said more than second logical reversing valves; The import of said more than first logical reversing valves is communicated with one of them the said reagent bottle that is used to hold damping fluid, washes said preparation pipeline and main pipe line.After a complete set of agent delivery that a reaction warehouse carries out single reaction finishes, wash said preparation pipeline and main pipe line, for a complete set of reagent of next reaction warehouse supply is prepared.

In order to prevent the interference between the adjacent reagent, in the present embodiment, also between step S3 and S4, increase following step:

S31: make the outlet of said more than first logical reversing valves and air connect for some time; This time calculates according to the time situation; Select between the 0.5s-1.5s, the requirement that does not influence sequencing reaction with the adjacent reagent of the satisfied isolation of the length of formed gas column in preparing pipeline is as the criterion.

Following with reference to Figure 32 and combine Fig. 1-Figure 31 that the system that is used for the dna sequencing appearance of present embodiment is described.

As stated, the dna sequencing appearance in the present embodiment comprises a plurality of reaction warehouses; Be used to gather the CCD camera 110 of the optical signal that dna sequencing reaction produced in each reaction warehouse; Be used to support CCD camera 110 can the two dimension adjustment bracing or strutting arrangement; Bracing or strutting arrangement comprise the first servomotor (not shown) that driven CCD camera 110 switches between different reaction warehouses and driven CCD camera 110 near or away from second servomotor 326 of one of them reaction warehouse, the bracing or strutting arrangement upper edge near or be provided with a plurality of transmitters 38 that are used to detect the CCD camera position away from the direction of reaction warehouse; And the agent delivery assembly that reaction reagent and damping fluid are provided for each reaction warehouse; The agent delivery assembly comprises first peristaltic pump 701 that is used to extract sequencing reaction reagent, be used to extract second peristaltic pump 711 of damping fluid, the more than second logical reversing valves more than 712, the first that are used for more than first logical reversing valves 702 of selective reagents and are used for the selective reaction storehouse lead to reversing valves 702 and more than second and lead to reversing valves 712 all for by motor-driven rotary valve.Promptly rotate by the electric motor driving spool; One of them import of logical reversing valve 702 more than first is communicated with outlet; The reagent bottle that be connected with this import this moment just is communicated with the outlet of more than first logical reversing valves 702; When needs changed reaction reagent, the electric motor driving rotary spool rotated to import that institute hopes to open is got final product with exporting to be communicated with.Working process of logical reversing valve 712 more than second and more than first logical reversing valves 702 are similar.When different was the motor rotation, change was led to the position of the anastomosing outlet of import of reversing valves 712 with more than second, because an outlet of each reaction warehouse and more than second logical reversing valves 712 is communicated with, led to the reaction warehouse that reversing valves 712 are communicated with thereby change with more than second.

Shown in figure 32, the system of present embodiment comprises PLC (programmable logic controller); PLC is electrically connected with the startup of controlling first servomotor and second servomotor respectively, stops and turning direction with the unit of the unit of first servomotor and second servomotor; PLC respectively with state first peristaltic pump, second peristaltic pump is electrically connected with the startup of controlling first peristaltic pump and second peristaltic pump, stops and rotating speed; PLC is electrically connected to control the on position of logical reversing valve more than first and more than second logical reversing valves with the drive-motor of more than first logical reversing valves and the drive-motor of more than second logical reversing valves respectively: the on position of PLC control more than first logical reversing valves 702 is to select reagent bottle anastomosing with it, and the on position of PLC control more than second logical reversing valves 712 is to select reaction warehouse anastomosing with it.PLC is electrically connected the open and close state with control CCD camera with the CCD camera; PLC also connects the position signal of the CCD camera that sends with receiving sensor respectively with a plurality of transmitters (transmitter only is shown, and other transmitters all are connected with PLC) in Figure 32.When CCD camera 110 is fitted on the reaction warehouse and reach the reaction times of expectation, open the CCD camera and obtain the optical signal that sequencing reaction produces, form image.

Shown in figure 32, the system of present embodiment also comprises the upper computer that is connected with said PLC, and said upper computer is used to be provided with the sequence of control of said PLC operation, and transmits said sequence of control to said PLC.Sequence of control is the workflow of PLC; That is to say PLC to the Be Controlled object, comprise drive-motor and the kind that CCD camera 110 transmits control signal and in proper order of drive-motor, the logical reversing valve 712 more than second of unit, first peristaltic pump 701, second peristaltic pump 711, the logical reversing valve 702 more than first of unit, second servomotor of first servomotor.In the present embodiment, upper computer carries out serial communication through RS232 port and PLC.Upper computer is communicated by letter with the CCD camera through USB port, is used to receive the view data that the CCD camera is obtained.

Following with reference to Figure 33, and combine Figure 11 that the temperature control modules of the system that is used for the dna sequencing appearance of embodiments of the invention is described.

Shown in figure 33, temperature control modules comprises the rly. that is connected with PLC and is arranged at TP 561, the semiconductor heat booster 571 in the reaction warehouse; PLC receives and is connected from the signal of TP 561 and with the primary return of rly., and the secondary loop of rly. is connected in the current supply circuit of semiconductor heat booster 571.When the temperature in the annular recesses 56 is lower than preset temperature; PLC sends connection signal to rly.; The current supply circuit of semiconductor heat booster 571 is connected; When the temperature in the annular recesses 56 is higher than another preset temperature; PLC sends signal to rly. breaks off the current supply circuit of semiconductor heat booster 571, guarantees that in view of the above the temperature that is positioned at annular recesses 56 is controlled at target temperature range.In the present embodiment, rly. is an electromagnetic relay.Only show the composition of the temperature control modules in the reaction warehouse among Figure 33, other reaction warehouses are identical therewith.Shown in figure 11; TP 561 is arranged in the reaction warehouse with semiconductor heat booster 571, is fixed with TP 561 in the bottom in sequencing reaction pond 51 through heat conductive silica gel, is not provided with the opposite side of annular recesses 56 at reaction tank body 5; Offer the heating chamber 57 that is used for ccontaining semiconductor heat booster 571; Heating chamber 57 is opened on the side of reaction tank body 5, and fixing semiconductor heat booster 571 on its bottom surface is with to 5 heating of reaction tank body.

In this enforcement, PLC adopts the CP1H-40XDT-D-CH type of Omron, controls first servomotor and second servomotor through pulse signal respectively, and first servomotor and second servomotor are stepper-motor; PLC controls the motor speed of first peristaltic pump and second peristaltic pump through sending pulse signal.Omron CP1H-40XDT-D-CH belongs to 40 PLC of rly. output type; 8 output expansions have been introduced simultaneously again; The BCD (binary coded decimal) that is used for more than first logical reversing valve and more than second logical reversing valves controls, through carrying out switching that BCD controls the on position of realizing valve etc.PLC makes the current supply circuit of semiconductor heat booster come controlled temperature through the break-make of relay.System of the present invention; The agent delivery assembly that makes the dna sequencing appearance with a plurality of reaction warehouses is for supply reagent and damping fluid timely and accurately; And can make the CCD camera in time read the optical signal in each reaction warehouse; Realized that a plurality of reactions carry out simultaneously, can check order to a plurality of samples simultaneously, improved dna sequencing efficient greatly.

Certainly, more than be preferred implementation of the present invention, should be understood that; For those skilled in the art; Under the prerequisite that does not break away from the principle of the invention, can also make some improvement and retouching, these improvement and retouching also are regarded as protection scope of the present invention.

Claims (5)

1. system that is used for the dna sequencing appearance, said dna sequencing appearance comprises a plurality of reaction warehouses; Be used to gather the CCD camera of the optical signal that dna sequencing reaction produced in each said reaction warehouse; Be used to support said CCD camera can the two dimension adjustment bracing or strutting arrangement; Said bracing or strutting arrangement comprise first servomotor that the driven CCD camera switches between different reaction warehouses with drive said CCD camera near or away from second servomotor of one of them reaction warehouse, said bracing or strutting arrangement upper edge near or be provided with a plurality of transmitters that are used to detect said CCD camera position away from the direction of said reaction warehouse; And be the agent delivery assembly that a plurality of said reaction warehouses provide reaction reagent and damping fluid; Said agent delivery assembly comprises first peristaltic pump, second peristaltic pump that is used to extract damping fluid that are used to extract sequencing reaction reagent, be used for more than first logical reversing valves of selective reagents and be used for the more than second logical reversing valves in selective reaction storehouse, and said more than first lead to reversing valves and more than second leads to reversing valves all for by motor-driven rotary valve;

It is characterized in that said system comprises: PLC;

Said PLC is electrically connected with the startup of controlling said first servomotor and second servomotor respectively, stops and turning direction with the unit of the unit of said first servomotor and second servomotor;

Said PLC respectively with state first peristaltic pump, second peristaltic pump is electrically connected with the startup of controlling said first peristaltic pump and second peristaltic pump, stops and rotating speed;

Said PLC is electrically connected to control the on position of logical reversing valve more than first and more than second logical reversing valves with the drive-motor of said more than first logical reversing valves and the drive-motor of more than second logical reversing valves respectively;

Said PLC is electrically connected with said CCD camera to control the open and close state of said CCD camera;

Said PLC also is connected with said a plurality of transmitters respectively to receive the position signal of the CCD camera that said transmitter sends.

2. system as claimed in claim 1; It is characterized in that; Also comprise temperature control modules; Said temperature control modules comprises the rly. that is connected with said PLC and is arranged at TP and the semiconductor heat booster in the said reaction warehouse, and said PLC receives and is connected from the signal of said TP and with the primary return of said rly., and the secondary loop of said rly. is connected in the current supply circuit of said semiconductor heat booster.

3. system as claimed in claim 1 is characterized in that, also comprises the upper computer that is connected with said PLC, and said upper computer is provided with the sequence of control of said PLC operation, and transmits said sequence of control to said PLC.

4. system as claimed in claim 3 is characterized in that, said upper computer is through RS232 port and said plc communication.

5. system as claimed in claim 3 is characterized in that, said upper computer is communicated by letter with said CCD camera through USB port, is used to receive the view data that said CCD camera is obtained.

Images