CN102604826A - Gene sequencing equipment - Google Patents
Gene sequencing equipment Download PDFInfo
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- CN102604826A CN102604826A CN2012100711644A CN201210071164A CN102604826A CN 102604826 A CN102604826 A CN 102604826A CN 2012100711644 A CN2012100711644 A CN 2012100711644A CN 201210071164 A CN201210071164 A CN 201210071164A CN 102604826 A CN102604826 A CN 102604826A
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Abstract
The invention relates to the field of biochemical equipment and provides gene sequencing equipment, which comprises a sequencing reaction chamber and a temperature control component. The sequencing reaction chamber is used for fixing samples and performing sequencing reaction; the temperature control component is fixed in an area of the sequencing reaction chamber when the samples are fixed and used for controlling temperature of the sequencing reaction chamber. The temperature of the sequencing reaction chamber rises and decreases quickly by the temperature control component, and sequencing reaction efficiency is improved.
Description
Technical field
The present invention relates to the biochemical equipment field, more particularly, relate to a kind of gene sequencing equipment.
Background technology
The blank of gene sequencing technology is formed at twentieth century, and existing nowadays should technology have developed like a raging fire is about to become a new technology that promotes progress of human society this century.The gene sequencing technology is to detect nucleic acid through gene sequencing equipment, thereby obtains the sequence of nucleic acid through data processing.
In the prior art, gene sequencing equipment comprises sequencing reaction small chamber, light source assembly, temperature control assembly, liquid transmission assembly, adopts figure assembly and leveling assembly.Wherein, said light source assembly comprises bright field light source and excitation light source.In this technical scheme, the liquid transmission assembly is used for storage and fluid transfer; Sequencing reaction small chamber is used for fixing sample and carries out sequencing reaction; The temperature control assembly is used to control the temperature of sequencing reaction small chamber; The leveling assembly is used to regulate the position of sequencing reaction small chamber; Adopt the image that the figure assembly is used to gather sequencing reaction.When carrying out sequencing reaction, the liquid transmission assembly is transferred to reagent in the sequencing reaction small chamber, carries out sequencing reaction; When sequencing reaction need heat up; The temperature control assembly adds thermal control to sequencing reaction small chamber, and when needs were lowered the temperature, sequencing reaction small chamber naturally cooled to specified temperature; When sequencing reaction was accomplished, the leveling assembly was regulated the position of sequencing reaction small chamber; Then, utilize bright field light source irradiation sequencing reaction small chamber, focus, make and adopt the figure assembly and can photograph distinct image, adopt the light field figure that the figure assembly is taken sequencing reaction in the sequencing reaction small chamber; Close bright field light source, utilize excitation light source irradiation sequencing reaction small chamber, focus, make and adopt the figure assembly and can photograph picture clearly, adopt the fluorogram that the figure assembly is taken sequencing reaction in the sequencing reaction small chamber.In this technical scheme, because light field lamp and excitation light source lay respectively at the both sides of sequencing reaction small chamber, thus require the regional have no occluder of sequencing reaction small chamber fixed sample, so that light field lamp and exciting light can shine the zone of sequencing reaction small chamber fixed sample.Therefore, the temperature control assembly can not directly carry out temperature regulation to the zone of sequencing reaction small chamber fixed sample.In the prior art; The temperature regulation to sequencing reaction small chamber is realized at the edge in the zone of temperature control assembly through being close to the sequencing reaction small chamber fixed sample; Because it is limited to be used for fixing the space of temperature control assembly; Simultaneously, the contact area of temperature control assembly and sequencing reaction small chamber is limited, causes the temperature control assembly can only be the less temperature control assembly that only possesses heating function of volume.Simultaneously, because the temperature control assembly is fixed in the edge of sequencing reaction small chamber, make that the edge of sequencing reaction small chamber can not fixed sample, the maximum area that causes being used for fixing in the sequencing reaction small chamber zone of sample is 1600mm
2Simultaneously, because existing bright field light source in the light source assembly has excitation light source again, thereby make the complex structure of gene sequencing equipment.
In the technique scheme, the temperature control assembly of gene sequencing equipment can not be realized heating and refrigerating function simultaneously, and limited owing to the fixed position, makes the intensification inefficiency.Simultaneously, because the structure of light source assembly, make the complex structure of gene sequencing equipment.In addition, because the cause of light source assembly and temperature control assembly, it is limited to make that sequencing reaction small chamber is used for fixing the area of sample area, thereby makes sequencing throughput not high.
Therefore, need a kind of gene sequencing equipment, can realize heating and refrigeration simultaneously, and the high function of the efficient of heating and cooling.
Summary of the invention
The object of the present invention is to provide a kind of gene sequencing equipment, be intended to solve prior art and can not realize heating and refrigeration simultaneously, and problem such as the efficient of heating and cooling is low.
In order to realize goal of the invention, said gene sequencing equipment comprises: sequencing reaction small chamber, temperature control assembly; Said sequencing reaction small chamber is used for fixing sample, and carries out sequencing reaction; Said temperature control assembly is fixed in the zone of sequencing reaction small chamber fixed sample, is used for sequencing reaction small chamber is carried out temperature control.
Wherein, said gene sequencing equipment also comprises light source assembly, is used to provide the light source of taking light field figure and the excitation light source of taking fluorogram; Said light source assembly also comprises light source, is positioned at a sequencing reaction small chamber side relative with the temperature control assembly, is used to the light that provides photographic images required.
Wherein, said light source assembly also comprises reflecting piece and filtering apparatus; Said reflecting piece between temperature control assembly and sequencing reaction small chamber, is used to reflect the light that arrives on the reflecting piece, takes the required light of light field figure to provide; Said filtering apparatus is positioned at a side of light source, is used to filter stray light.
Further, said filtering apparatus also is used to change the light path through the light of said filtering apparatus.
Wherein, the thermal conductivity of said reflecting piece is at least 80W/m.k, and its surface is at least 60% material by reflecting rate and processes.
Wherein, said gene sequencing equipment also comprises the liquid transmission assembly, is used for storage and transmission reagent.
Wherein, said liquid transmission assembly comprises device for storing liquid and passes liquid device; Said device for storing liquid is used to store at least a reagent; Said biography liquid device is positioned at a side of device for storing liquid, is used for the reagent of device for storing liquid is transferred to sequencing reaction small chamber.
Wherein, said temperature control assembly comprises temperature measuring equipment, DHE, cooling piece and radiator element.Said temperature measuring equipment has TP, and TP is used for measuring temperature.Said temperature measuring equipment is connected with sequencing reaction small chamber, is used for the temperature through the temperature sensor measurement sequencing reaction small chamber, and the temperature of measuring is sent to DHE; Said DHE is connected with temperature measuring equipment, is used to receive and handle the temperature that temperature measuring equipment sends, and produces steering order; Said cooling piece is connected respectively with sequencing reaction small chamber with DHE, is used for carrying out work according to steering order, so that the sequencing reaction small chamber heating or the required energy that freezes to be provided; Said radiator element is connected with cooling piece, is used to transmit the energy of cooling piece one side.
Wherein, said temperature control assembly comprises temperature measuring equipment, DHE, heating piece and refrigeration plant.Said temperature measuring equipment comprises TP.Said temperature measuring equipment is connected with sequencing reaction small chamber, is used for the temperature through the temperature sensor measurement sequencing reaction small chamber, and the temperature of measuring is sent to DHE; Said DHE is connected with temperature measuring equipment, is used to receive and handle the temperature that temperature measuring equipment sends, and produces steering order; Said heating piece is connected respectively with DHE with sequencing reaction small chamber, is used for carrying out work according to steering order, to provide the sequencing reaction small chamber heating required energy; Said refrigeration plant is positioned at liquid transmission assembly below, is used for the reagent in the liquid transmission assembly is freezed, and refrigerating reagent is transferred to sequencing reaction small chamber through the liquid transmission assembly, so that sequencing reaction small chamber is freezed.
Wherein, the area that is used for fixing the zone of sample in the said sequencing reaction small chamber is S; Preferably, 150mm
2≤S≤2100mm
2
Wherein, said sequencing reaction small chamber is hyperchannel or single passage sequencing reaction small chamber.
By on can know that in the gene sequencing equipment of the present invention, the temperature control assembly can heat and can freeze sequencing reaction small chamber.Simultaneously, the temperature control assembly fully contacts with the zone of sequencing reaction small chamber fixed sample, makes that the transmission ofenergy between them is more rapid, thereby has improved warming and cooling rate, and then improved the efficient of order-checking.In addition, but the equal fixed sample in optional position in the sequencing reaction small chamber has avoided the sequencing reaction small chamber edge to be used for fixing the temperature control assembly and can not fixed sample, and this scheme makes that the zone of sequencing reaction small chamber fixed sample is bigger, thereby has improved sequencing throughput.And gene sequencer of the present invention also can provide photographic images required light through same light source, makes that the gene sequencing device structure is simple.
Description of drawings
Fig. 1 is the structural representation of gene sequencing equipment in the one embodiment of the invention;
Fig. 2 is the structural representation of sequencing reaction small chamber in one embodiment of the invention;
Fig. 3 is the structural representation of sequencing reaction small chamber in another embodiment of the present invention;
Fig. 4 is the structural representation of gene sequencing equipment in the another embodiment of the present invention;
Fig. 5 is the structural representation of temperature control assembly in the another embodiment of the present invention;
Fig. 6 is the structural representation of light source assembly in the one embodiment of the invention;
Fig. 7 is the structural representation of light source assembly in the another embodiment of the present invention;
Fig. 8 is the structural representation of spectral filter in the one embodiment of the invention;
Fig. 9 is the structural representation of spectral filter in the one embodiment of the invention;
Figure 10 is the structural representation of gene sequencing equipment in the another embodiment of the present invention;
Figure 11 is the explosive view of temperature control assembly in the one embodiment of the invention;
Figure 12 is a liquid transmission assembly structural representation in the one embodiment of the invention;
Figure 13 is the structural representation of gene sequencing equipment in the another embodiment of the present invention.
Embodiment
In order to make the object of the invention, technical scheme and advantage clearer,, the present invention is further elaborated below in conjunction with accompanying drawing and embodiment.
The sample and the reagent of the sequencing reaction small chamber internal fixing among the present invention react, and make sample carry affinity tag, and form the sample that carries affinity tag, can be luminous after this sample that carries affinity tag passes through excitation.
Sample among the present invention is through globule or directly be fixed on the sequencing reaction small chamber, and said sample is fixed on (this process is point sample) on the sequencing reaction small chamber, and the formed point one by one of sample after fixing is called array point.Density one timing of array point in sequencing reaction small chamber, the zone that is used for fixing sample in the sequencing reaction small chamber is big more, and the quantity of fixed array point is many more, also is that sequencing throughput is high more.
To gene sequencing equipment, the present invention proposes an embodiment, and is of Fig. 1, and gene sequencing equipment comprises sequencing reaction small chamber and temperature control assembly.Wherein:
(1) sequencing reaction small chamber 1, is used for fixing sample 10, and carries out sequencing reaction.
Said sequencing reaction small chamber is used for fixing sample, and sequencing reaction takes place under the control of temperature control assembly for sample and reagent.Said sequencing reaction small chamber does not have concrete restriction.Preferably; Sequencing reaction small chamber is the sequencing reaction small chamber of three-decker; The first layer is used for fixing sample, and the second layer is used for forming the groove of hollow, the 3rd layer of groove that is used for forming with the first layer and the second layer hollow of sealing; The two ends perforate of the groove of the middle control of sealing is used for supplying reagent to flow into and outflow.Preferably, sequencing reaction small chamber is a bilayer structure, and the first layer is used for fixing sample, and the second layer is used for forming with the first layer the groove of the hollow of sealing, two STHs of said sequencing reaction small chamber, thus is communicated with the groove of hollow, form the reagent inlet and export.
(2) the temperature control assembly 2, are fixed in the zone of sequencing reaction small chamber 1 fixed sample 10, are used for sequencing reaction small chamber 1 is carried out temperature control.
Said temperature control assembly can carry out temperature survey for having, and can realize heating and refrigerating device, thereby realizes the sequencing reaction small chamber accurate temperature controlling.Also promptly this temperature control assembly has temperature measuring equipment and is used for heating and refrigerating device, through thermometric temperature measurement device, then through heating and the refrigerating device comes sequencing reaction small chamber is heated or freezes.
In the present embodiment, sample 10 is fixed in the sequencing reaction small chamber 1, and temperature control assembly 2 is fixed on the zone of sequencing reaction small chamber 1 fixed sample 10.When sequencing reaction small chamber 1 needs heating or refrigeration, 2 pairs of sequencing reaction small chamber heating of temperature control assembly or refrigeration.In the present embodiment, sequencing reaction small chamber 1 is not limit with the area size of the contact surface of temperature control assembly 2.Preferably, temperature control assembly 2 is used to heat or the area size with sequencing reaction small chamber fixed sample 10 is the same at least in the zone of refrigerating parts.Further preferred, temperature control assembly 2 is used to heat or the area of the face that the area of refrigerating parts and sequencing reaction small chamber 1 contact with temperature control assembly 2 is identical, also is that sequencing reaction small chamber 1 and temperature control assembly 2 just can overlap.
In the present embodiment, the zone of fixed sample is fixed with sample 10, and temperature control assembly 2 directly or indirectly contacts with the zone of sequencing reaction small chamber 1 fixed sample, makes that the warming and cooling rate of gene sequencing equipment is faster, thereby has improved the efficient of order-checking.Simultaneously; Because the size of temperature control assembly 2 does not receive space constraint; Can heat also can refrigerating temperature control assembly 2 so this temperature control assembly 2 can have enough spaces to place; Prevented that temperature control assembly 2 is connected with the sequencing reaction small chamber lateral edges, thereby and caused sequencing reaction small chamber both sides gravity unbalance and make that sequencing reaction small chamber is unbalance to make sequencing reaction small chamber in adopting the figure process, focus error to occur.In addition; The area of the area of the face of temperature control assembly and sequencing reaction small chamber contact and this face of sequencing reaction small chamber is consistent; Also be contact surface big or small in full accord of temperature control assembly and sequencing reaction small chamber; Temperature control assembly 2 be used for the heating or the refrigerating region area big, thereby make that sequencing reaction small chamber 2 warming and cooling rates are faster.In addition, relative prior art, the former zone that is used for fixing the temperature control assembly of sequencing reaction small chamber 1 also can be used for fixed sample in the present technique scheme, and the zone of sequencing reaction small chamber internal fixing sample increases, thereby makes that the sequencing throughput of gene sequencing equipment is higher.
Based on a last embodiment, the present invention proposes another embodiment, and is as shown in Figure 1.Said gene sequencing equipment comprises sequencing reaction small chamber, temperature control assembly, light source assembly.Wherein:
(1) sequencing reaction small chamber 1, is used for fixing sample 10, and carries out sequencing reaction.
(2) the temperature control assembly 2, are fixed in the zone of sequencing reaction small chamber 1 fixed sample 10, are used for sequencing reaction is carried out temperature control.
(3) light source assembly 3, are used to provide the light source of taking light field figure and the excitation light source of taking fluorogram.
Said light source assembly comprises light source, is positioned at a sequencing reaction small chamber side relative with the temperature control assembly, is used to the light that provides photographic images required.
The sequencing reaction small chamber 1 internal fixing sample 10 of the gene sequencing equipment in the present technique scheme, 2 pairs of sequencing reaction small chambers of temperature control assembly 1 carry out temperature control, and sequencing reaction can be carried out smoothly.When the image in the needs shooting sequencing reaction small chamber 1, light source assembly 3 provides photographic images required light, and light arrives sequencing reaction small chamber 1, for photographic images.The position of the light source assembly 3 of this gene sequencing equipment is specifically restriction not, makes that temperature control assembly 2 can be bigger with the contact area of sequencing reaction small chamber 1 fixed sample, thereby has improved heating or refrigerating efficient greatly.Simultaneously; Because it is limited to be used for fixing the area size of temperature control assembly 2 in traditional gene sequencing equipment; Can fixed the zone can only carry temperature control assembly 2 with heating function; And fixedly the zone of temperature control assembly 2 is not limited in the technical scheme of the present invention, thereby makes temperature control assembly 2 of the present invention can realize that heating can realize refrigeration again.In addition, because technical scheme light source assembly of the present invention is simple in structure, make that the gene sequencing device structure is simple.
The area that is used for fixing the zone of sample in the sequencing reaction small chamber among the present invention is S, and S is any.Preferably, 150mm
2≤S≤2100mm
2Wherein, how many selections of the area of sequencing reaction small chamber size is selected according to required fixed sample, needs the fixed sample many, then selects the big sequencing reaction small chamber of area, needs the fixed sample few, then selects the little sequencing reaction small chamber of area.Sequencing reaction small chamber of the present invention can be changed as required, when guaranteeing sequencing throughput, has practiced thrift the reagent of order-checking, has also promptly practiced thrift the cost of order-checking.Under the identical situation of array point density, sequencing reaction small chamber of the present invention can fixed sample zone maximum area be 2100mm
2Thereby, improved the sequencing throughput of gene sequencing equipment.
To sequencing throughput, the present invention proposes an embodiment, and the area that in sequencing reaction small chamber, is used for fixing sample area is 2000mm
2, the density of array point is 1 * 10
5Individual/mm
2Then the number of fixable array point can reach 2 * 10 in this sequencing reaction small chamber
8, also be that the order-checking passage of gene sequencing equipment reaches 2 * 10
8Gene sequencing equipment in the present embodiment has been realized the high-throughput gene sequencing.Wherein, the density of array point is the flux density of coming setting array point according to order-checking, needs flux high and be not very high to the specification of quality of the image of follow-up bat figure, and then the density of array point can height, even can reach 1 * 10
6Individual/mm
2Otherwise, then can reduce the density of array point.
To sequencing reaction small chamber, the present invention proposes an embodiment, and is as shown in Figure 2, and sequencing reaction small chamber is the single passage sequencing reaction small chamber.Sample is fixed in the reaction channel 11 of sequencing reaction small chamber 1, and during reaction, reagent flows into from an end of reaction channel 11; Flood sample, flow out from the other end then, make sample and reagent full contact; React more complete, and make sequencing reaction in having the reagent of diffusion barrier, not carry out.The high-flux sequence of single sample is realized in this zone.
To sequencing reaction small chamber, the present invention proposes another embodiment, and is as shown in Figure 3, and sequencing reaction small chamber is the hyperchannel sequencing reaction small chamber.Sample is fixed in the reaction channel 11 of sequencing reaction small chamber 1, and during reaction, reagent flows into from an end of reaction channel 11, flows out from the other end, and when reagent flow through reaction channel 11, reagent flooded sample, made sample and reagent fully react.Simultaneously, reagent flows to the other end from an end of reaction channel 11, makes sequencing reaction in having the reagent of diffusion barrier, not carry out.Wherein, the number of reaction channel 11 is preferably between 2～16, such as 4 reaction channels, 8 reaction channels etc.The number of can number per sample coming selective reaction passage 11, the sample that also promptly need survey more (comprise kind is more, single sample quantity more), the reaction channel 11 of the sequencing reaction small chamber 1 of selection is many more.Preferably, a kind of sample of each reaction channel 11 internal fixing also is all to be fixed with sample on the reaction channel of sequencing reaction small chamber 1; There is not empty reaction channel 11; The sample of each reaction channel 11 internal fixing can be identical, also can be different, such as 12 reaction channels, 11 fixing eight kinds of different samples; 6 fixing six kinds of different samples of reaction channel, 16 reaction channel 11 fixing a kind of samples.The area S that is used for fixing the zone of sample in the sequencing reaction small chamber 1 is preferably 150mm
2～1100mm
2, the area S0 of each reaction channel 11 is preferably 100mm
2～1000mm
2How many areas of said reaction channel 11 is selected according to fixed sample.
Based on the foregoing description, the present invention proposes another embodiment, and is as shown in Figure 4, and this gene sequencing equipment comprises liquid transmission assembly, sequencing reaction small chamber, temperature control assembly, light source assembly, adopts the figure assembly.Wherein:
(1) sequencing reaction small chamber 1, is used for fixing sample, and carries out sequencing reaction.
(2) the temperature control assembly 2, are fixed in the zone of sequencing reaction small chamber fixed sample, are used for sequencing reaction small chamber is carried out temperature control.
(3) light source assembly 3, are used to provide the light source of taking light field figure and the excitation light source of taking fluorogram.
(4) the liquid transmission assembly 4, are used for storage, transmission reagent.
Said liquid transmission assembly comprises the device that is used to transmit reagent; This device can be in combination of mechanical manipulator, pump, pump and valve etc. any one; The concrete structure of liquid transmission assembly waits according to liquid transmitting speed, precision and makes up, and the liquid transmission assembly in the present embodiment does not have concrete restriction.Wherein this liquid transmission assembly can comprise that also the reagent platform is used for storage of liquids.
(5) adopt figure assembly 5, be used for photographic images.
The said figure of adopting assembly comprises the image sensor of clapping map device.Preferably, this adopt the figure assembly comprise charge coupled cell (Charge-coupled Device, CCD), cmos camera.Wherein adopt the figure assembly and also can comprise gearshift, be used for the position relation of image sensor and sequencing reaction small chamber, to guarantee to photograph distinct image.
In the present technique scheme, sequencing reaction small chamber 1 fixed sample of gene sequencing equipment, liquid transmission assembly 4 is transferred to sequencing reaction small chamber 1 with liquid; 2 pairs of sequencing reaction small chambers of temperature control assembly 1 carry out temperature control; Sample and reagent carry out sequencing reaction in sequencing reaction small chamber 1, when the needs photographic images, light source assembly 3 provides photographic images required light; Light arrives sequencing reaction small chamber 1, adopts the sequencer map picture that figure assembly 5 is taken in the sequencing reaction small chamber.In the technical scheme of present embodiment, the gene sequencing device structure is simple, and temperature control assembly 2 fully contacts with sequencing reaction small chamber 1, makes that temperature control efficient is higher.
To the liquid transmission assembly, the present invention proposes an embodiment, and is as shown in Figure 5.The liquid transmission assembly comprises device for storing liquid and passes liquid device.Wherein:
(1) device for storing liquid 41, are used to store at least a reagent.
(2) pass liquid device 42, be positioned at a side of device for storing liquid 41, be used for the reagent of device for storing liquid 41 is transferred to sequencing reaction small chamber.
Device for storing liquid 41 in the present embodiment can store different reagent, and this device for storing liquid 41 can be changed according to kind that holds reagent and dosage.Pass liquid device 42 reagent in the device for storing liquid 41 is transferred to sequencing reaction small chamber, pass liquid device 42 and can be among pump, valve, the mechanical manipulator three at least one.How much pass liquid device 42 can choosing according to the amount of the speed of the speed that extracts liquid, fluid transfer, fluid transfer.
Technical scheme in the present embodiment can be changed device for storing liquid and pass liquid device according to different demands, makes that the suitability of liquid transmission assembly is good.
Fig. 6 shows the structure of light source assembly in one embodiment of the invention.Light source assembly comprises light source and reflecting piece.Wherein:
(1) light source 31, are positioned at the relative side of sequencing reaction small chamber 1 and temperature control assembly 2, are used to the light that provides photographic images required.
(2) reflecting piece 32, between temperature control group 2 and sequencing reaction small chamber 1, are used to reflect the light that arrives on the reflecting piece, take the required light of light field figure to provide.
(3) filtering apparatus 33, are positioned at a side of light source 31, are used to filter stray light.
In the present embodiment, light source 31 carries affinity tag sample luminous light source for exciting arbitrarily.Be preferably the laser equipment that mercury lamp maybe can send laser.This light source assembly has a light source 31 at least, all is positioned at the same side of sequencing reaction small chamber.When light source 31 when being a plurality of, each light source 31 can send the light of specific wavelength, and the light of different wave length can excite the sample that carries affinity tag in the sequencing reaction small chamber 1 to send the light of distinct colors; When gathering fluorogram, open certain light source 31, move this light source 31; The luminous energy that makes light source send gets at and reaches sequencing reaction small chamber, and it is luminous that the sample of affinity tag is carried in the optical excitation of this specific wavelength, simultaneously; The light of specific wavelength is by reflecting piece 32 reflection, carries light that the sample of affinity tag sends and reflected light through filtering apparatus 33, filtering apparatus 33 filtration reflected light; The figure assembly is adopted in light arrival through filtering apparatus 33, adopts figure assembly photographic images.When the light time of the other a kind of specific wavelength of needs, open a kind of light source 31 that can send the light of this specific wavelength in addition, adjust the position of this light source 31; The luminous energy that makes light source send gets at and reaches sequencing reaction small chamber; It is luminous that the sample of affinity tag is carried in the optical excitation of this specific wavelength, and simultaneously, the light of specific wavelength is by reflecting piece 32 reflections; Carry light that the sample of affinity tag sends and reflected light through filtering apparatus 33; Filtering apparatus 33 filters reflected light, adopts the figure assembly through the light arrival of filtering apparatus 33, adopts figure assembly photographic images.Switch and open the light source 31 of the light that can send different wave length as required, till photographing required all fluorograms.When gathering light field figure, open a plurality of light sources 31 simultaneously, make light arrive reflecting piece 32; And after after reflecting piece 32 reflections, (according to the needs of clapping figure, filtering apparatus 33 allows the light of certain section wavelength in this mixed light to pass through to filter stray light through filtering apparatus 33; Other light are by filtering); Move and to adopt the figure assembly, light after the filtration is arrived adopt the figure assembly, adopt the figure assembly and gather image.
When light source assembly had only a light source 31, light source 31 can be sent out the light of different wave length.When needs are taken light field figure; Light is injected sequencing reaction small chamber 1 and is arrived on the reflecting piece 32, and reflecting piece 32 reflections arrive the light of reflecting piece 32, and this light is through filtering apparatus 33; Filtering apparatus 33 filters stray light (according to the requirement of clapping figure, filtering apparatus allows the light of certain section specific wavelength to pass through); When needs were taken fluorogram, light was through filtering apparatus 33, and the light of specific wavelength is through filtering apparatus 33; Arrive sequencing reaction small chamber 1, excite the sample that carries affinity tag in the sequencing reaction small chamber 1 luminous, the light of specific wavelength is reflected by reflecting piece simultaneously; This reflected light is luminous through filtering apparatus 33 with the sample that carries affinity tag; This reflected light is carried luminous the arrival through filtering apparatus 33 of sample of affinity tag and adopts the figure assembly by filtering, adopts the figure assembly and takes fluorogram.
In the present technique scheme, light source is positioned at the same side of sequencing reaction small chamber, makes the simple in structure of this gene sequencing equipment.Simultaneously; Avoided in the prior art scheme light source to be positioned at both sides and made sequencing reaction small chamber fixed sample zone have no occluder, thereby caused the temperature control assembly to be positioned on the edge of sequencing reaction small chamber, and made the temperature control inefficiency; But the zone of present technique scheme sequencing reaction small chamber fixed sample is bigger; So flux is higher, and the temperature control assembly is fixed in the zone at sequencing reaction small chamber fixed sample place, thereby improved heating or refrigerating efficient.
Among the present invention, the preferred thermal conductivity of reflecting piece is at least 80W/m.k, and process for reflecting rate is at least 60% material on its surface.This material can be solid or liquid.When being liquid, this liquid through transparent solid interlayer on one side encapsulate flap.When being solid, can directly make ganoid with this solid matter, when being pulverulent solids, can this pulverulent solids be spread upon the flap surface, make it have slick surface through handling.
Preferably; This reflecting piece is: the sheet that chrome faced gold and silver, copper, aluminium or iron are processed; The sheet of perhaps processing for the alloy of chrome faced gold and silver, copper, aluminium or iron and any metal; The sheet of perhaps processing for the gold of electroplate, copper, aluminium or iron, the sheet of perhaps processing, the perhaps sheet processed of stainless steel for chrome faced gold, copper, aluminium or iron and the alloy of any metal.Reflecting piece among the present invention is between sequencing reaction small chamber and temperature control assembly; This reflecting piece has good reflecting effect and heat-conducting effect; Thereby make the temperature control assembly heat evenly and fast passed to sequencing reaction small chamber, thereby improve the heating or the refrigerating speed of temperature control assembly greatly through reflecting piece.The good reflector efficiency of this reflecting piece simultaneously makes the luminous energy major part that arrives reflecting piece reflex to and is used for adopting the figure assembly, adopts the figure assembly and photographs distinct image thereby make.
The size of the reflecting piece among the present invention is big or small identical with the zone of sequencing reaction small chamber internal fixing sample at least.Guarantee to arrive the light of sequencing reaction small chamber, after the reflecting piece reflection, can illuminate the All Ranges of sequencing reaction small chamber fixed sample, thereby make and adopt the All Ranges that the figure assembly can clearly photograph the sequencing reaction small chamber fixed sample.
To light source assembly, the present invention proposes another embodiment, and is as shown in Figure 7.Shown light source assembly comprises light source, filtering apparatus and reflecting piece.Wherein:
(1) light source 31, are positioned at a sequencing reaction small chamber side relative with the temperature control assembly, are used to the light that provides photographic images required.
(2) reflecting piece 32, between temperature control assembly and sequencing reaction small chamber, are used to reflect the light that arrives on the reflecting piece, take the required light of light field figure to provide.
(3) filtering apparatus 33, are positioned at a side of light source, are used to filter stray light, and are used to change the light path through the light of said filtering apparatus.
In the present embodiment, each filtering apparatus 33 comprises many group spectral filters 330, and every group of spectral filter 330 allows the light of the specific wavelength in the light that light sources 31 send to pass through.When the light time that needs different wave length, rotate filtering apparatus 33 and to the spectral filter 330 of correspondence, get final product.Filtering apparatus 33 in this enforcement is used to change the light path through the light of filtering apparatus, and what make enough light arrive to be used for photographic images adopts the figure assembly.Also promptly as shown in Figure 8, light that light source sends and the light that from filtering apparatus 33, comes out is not on same straight line or plane.In the present embodiment, light source carries affinity tag sample luminous light source for exciting arbitrarily.Be preferably the laser equipment that mercury lamp maybe can send laser.
In the present embodiment, when taking light field figure, the light that light source 31 sends; Through behind the filtering apparatus 33, filtering apparatus 33 changes the light path of light, and light arrives reflecting piece 32 through sequencing reaction small chamber 1; This light excites also that to carry the affinity tag sample luminous; Reflecting piece 32 reflected light carry that the affinity tag sample is luminous to filter and change light path through filtering apparatus 33 with reflected light, make light arrive certain position that needs light (also promptly arrive be used for photographic images adopt the figure assembly).When taking fluorogram, the light that light source 31 sends is through behind the filtering apparatus 33; Filtering apparatus 33 filters stray lights, and the light after the filtration is reflected in the sequencing reaction small chamber 1, and the light after the sample that carries affinity tag in the sequencing reaction small chamber 1 is filtered excites; Send fluorescence, fluorescence is through filtering apparatus 33, and filtering apparatus 33 filters stray light; And the change light path, the light after make filtering arrive be used for photographic images adopt the figure assembly, thereby realized that light source 31 provides bat figure required light source; Filtering apparatus 33 filters stray light and changes light path, and reflecting piece 32 reflections arrive the light on the reflecting piece.The technical scheme of present embodiment; Light source both provided the light source of taking light field figure that the light source of taking fluorogram also is provided, and made that the structure of gene sequencing equipment is simpler, simultaneously; This filtering apparatus not only can filter stray light; And can change light path, and to adopt in the figure assembly thereby make required competent light arrive, final assurance is adopted the figure assembly and is photographed distinct image.
To spectral filter, the present invention proposes an embodiment, and is as shown in Figure 8.This spectral filter comprises two spectral filters, is respectively spectral filter 3301 and spectral filter 3302.The light of the specific wavelength in the light that spectral filter 3301 permission light sources send passes through; The light of specific wavelength is through behind the spectral filter 3301; The light of spectral filter 3302 these specific wavelengths of reflection also is that spectral filter 3301 filters stray light, the light path of the light that spectral filter 3302 change light sources send.The long light of said spectral filter 3302 reflection specific wavelengths allows the light of other wavelength to pass through.
In the technical scheme of present embodiment; Spectral filter has realized that successfully the light of specific wavelength arrives sequencing reaction small chamber; It is luminous that thereby the sample of affinity tag is carried in the optical excitation of specific wavelength; The sample that carries affinity tag is luminous, thereby makes the bat map device of adopting in the figure assembly can photograph the image in the sequencing reaction small chamber.Wherein, Being used for taking interior fluorogram of sequencing reaction small chamber and the bat map device of light field figure in the described figure of the adopting assembly does not limit; This device is preferably CCD (Charge-coupled Device; Charge coupled cell) or CID (charge injection device, charge-injection detector(CID)) or cmos camera.
To spectral filter, the present invention proposes another embodiment, and is as shown in Figure 9.This spectral filter comprises three spectral filters, is respectively spectral filter 3301, spectral filter 3302 and spectral filter 3303.The light of the specific wavelength in the light that spectral filter 3301 permission light sources send passes through; The light of specific wavelength is through behind the spectral filter 3301; Spectral filter 3302 changes the light path of the light of this specific wavelength; The light of specific wavelength arrives sequencing reaction small chamber through spectral filter 3303, also is that spectral filter 3301 filters stray light, the light path of the light that spectral filter 3302 change light sources send.The light of said spectral filter 3302 reflection specific wavelengths allows the light of other wavelength to pass through, and said spectral filter 3303 allows the light of certain section wavelength to pass through.
In the present embodiment, spectral filter can filter stray light, allows the light of specific wavelength to pass through, and the light of this specific wavelength can be the light of one section wavelength or certain concrete wavelength.After the light of specific wavelength arrives sequencing reaction small chamber; The sample that carries affinity tag in the optical excitation sequencing reaction small chamber of specific wavelength is luminous; Carry light that the sample of affinity tag sends through spectral filter, spectral filter filters stray light once more, makes the light that needs arrive and adopts the figure assembly.The bat map device of adopting in the figure assembly in the present embodiment is that any optical signalling converts analog current signal into, and current signal is realized the bat map device that obtains, stores, transmits, handles and reappear of image through amplifying and analog.Be preferably CCD or CID.
In the technical scheme of present embodiment, the stray light in the light that spectral filter filtration light source sends has realized that successfully the light of specific wavelength arrives sequencing reaction small chamber, and light arrives and adopts the figure assembly through after reflection or exciting, and photographs distinct image so that adopt the figure assembly.And this spectral filter can change light path, makes that the installation site of clapping the figure assembly is relatively freer, as long as be positioned at the same side with light source assembly.
Figure 10 shows the structure of gene sequencing equipment.Reflecting piece 32 is between temperature control assembly 2 and sequencing reaction small chamber 1.The light that light source 31 sends is through filtering apparatus 33, and filtering apparatus 33 filters stray light, changes light path; Make illumination inject sequencing reaction small chamber 1, light arrives reflecting piece 32, reflecting piece 32 reflected light through sequencing reaction small chamber; Make light pass through filtering apparatus 33; The stray light that filtering apparatus 33 filters in the reflected light, remaining light arrives and adopts figure assembly 5, adopts the image that figure assembly 5 is taken in the sequencing reaction small chamber.In the present embodiment, vertically inject the zone that is fixed with sample of sequencing reaction small chamber 1 through the light of filtering apparatus 33 change light paths.Reflecting piece 32 is between sequencing reaction small chamber 1 and temperature control assembly 3.The minimum size of reflecting piece 32 big or small identical for the zone that is fixed with sample.Preferably, the size of reflecting piece 32 is the same with sequencing reaction small chamber 1 size.In the present embodiment, reflecting piece 32 has good thermal conductivity.The thickness h of reflecting piece 32 is not limit, and is preferred, h≤0.1mm.
The light that technical scheme in the present embodiment, light source are sent is after the filtering apparatus filtration; The exciting light that had both stayed the specific wavelength that needs arrives sequencing reaction small chamber, to excite the sample that carries affinity tag in the sequencing reaction small chamber luminous, adopts the figure assembly through arriving behind the filtering apparatus; Thereby make that adopting the figure assembly photographs fluorogram; Can make again arrive sequencing reaction small chamber through the reflecting piece reflection, light reflexed to adopts the figure assembly, make and adopt the figure assembly and photograph light field figure clearly.The technical scheme of present embodiment; Utilize a light source to realize the shooting of light field figure and fluorogram; Not only convenient; Also avoided sequencing reaction small chamber when taking light field figure and fluorogram, to focus respectively, caused the focusing of light field figure and fluorogram inconsistent, and then make follow-up data handle the inaccurate phenomenon of data of the nucleotide sequence that obtains.
Temperature control assembly among the present invention proposes an embodiment, and is shown in figure 11.The temperature control assembly comprises cooling piece, radiator element and temperature measuring equipment.Wherein:
(1) temperature measuring equipment 21, are connected with sequencing reaction small chamber 1, are used for the temperature through temperature sensor measurement sequencing reaction small chamber 1, and the temperature of measuring is sent to DHE 20.
Said TP is experiencing temperature arbitrarily and converting the TP of usable output signal to of selling on the market.Be preferably contact type temperature sensor.
(2) DHE 20 is connected with temperature measuring equipment 21, is used to receive and handle the temperature that temperature measuring equipment sends, and produces steering order.
Said DHE is can processing data, produces the device of instruction.Be preferably the DHE that any one and peripheral circuit in chip, the micro-chip are formed.Further be preferably Personal Computer.
(3) cooling piece 22, are connected respectively with sequencing reaction small chamber 1 with data handling component 20, are used for carrying out work according to steering order, to provide sequencing reaction small chamber 1 heating or the required energy that freezes.
Said cooling piece is the cooling piece of selling on the market arbitrarily.The number of cooling piece 22 is not limit, and is preferably 1～5.
(4) radiator element 23, are connected with cooling piece 22, are used to transmit the energy of cooling piece 22 1 sides.
Said radiator element is coated with the last layer heat-conducting silicone grease at cooling piece and radiator element contact surface, and the heat that cooling piece is sent is more effective to be transmitted on the radiator element, is dispersed in the ambient air through radiator element and goes.This radiator element is a duraluminum, and brass or bronze are made tabular, sheet, the device of splintery etc.In this no particular restriction, the radiator element of selling on the market is all in the present invention available.
In the present embodiment, temperature measuring equipment 21 is close to sequencing reaction small chamber 1.Wherein a side of sequencing reaction small chamber 1 is temperature measuring equipment 21 and cooling piece 22, and radiator element 23 is positioned at the relative side of cooling piece 22 and sequencing reaction small chamber 1.After cooling piece 22 feeds electric current; 22 pairs of sequencing reaction small chambers of cooling piece, 1 heating (or refrigeration); The energy of the side that cooling piece 22 is connected with radiator element 23 through radiator element 23 with the transmission ofenergy of cooling piece 22 these sides in ambient air; Thereby make the temperature of the side that cooling piece 22 is connected with radiator element 23 descend rapidly (or rising), thereby the temperature of cooling piece 22 opposite sides is risen (or decline) rapidly, thereby reach purpose sequencing reaction small chamber 1 rapid heating (or refrigeration).Temperature measuring equipment 21 is measured the temperature of sequencing reaction small chamber 1, and the temperature that records is sent to DHE 20 in real time, and DHE produces steering order according to the temperature that receives; Make cooling piece 22 carry out work, thereby realize heating (or refrigeration) sequencing reaction small chamber 1 according to steering order.Wherein, cooling piece 22 is connected with DHE 20 through lead with temperature measuring equipment 21.The number of the temperature measuring equipment in the present embodiment is selected as required, when the accurate feedback temperature of needs, a plurality of temperature measuring equipments can be arranged.TP is not limit in the present embodiment.
Technical scheme in the present embodiment, temperature control assembly have realized the rapid heating of sequencing reaction small chamber or refrigeration have been improved the efficient of sequencing reaction, make that simultaneously temperature control is more accurate, have improved the quality of sequencing reaction.
To the temperature control assembly, the present invention proposes another embodiment, and is shown in figure 12.The temperature control assembly comprises heating piece, temperature measuring equipment and refrigeration plant.Wherein:
(1) temperature measuring equipment 21 is connected with sequencing reaction small chamber 1, is used for the temperature through the temperature sensor measurement sequencing reaction small chamber, and the temperature of measuring is sent to DHE 20.
Said TP is experiencing temperature arbitrarily and converting the TP of usable output signal to of selling on the market.Be preferably contact type temperature sensor.
(2) DHE 20 is connected with temperature measuring equipment 21, is used to receive and handle the temperature that temperature measuring equipment sends, and produces steering order.
Said DHE is can processing data, produces the device of instruction.Be preferably the DHE that any one and peripheral circuit in chip, the micro-chip are formed.Further be preferably Personal Computer.
(3) heating piece 24, are connected respectively with DHE with sequencing reaction small chamber, are used for carrying out work according to steering order, to provide the sequencing reaction small chamber heating required energy.
Said heating piece does not have particular restriction, is preferably electric heating sheets or silica gel heating piece, and it can the evolution of heat under the energising situation, thereby realizes the heating to sequencing reaction small chamber.
(4) refrigeration plant 25, are positioned at liquid transmission assembly below, are used for the reagent in the liquid transmission assembly is freezed, and refrigerating reagent is transferred to sequencing reaction small chamber through the liquid transmission assembly, so that sequencing reaction small chamber is freezed.
Said refrigeration plant be for being reduced to the device below 10 degrees centigrade with temperature, other no particular restriction.Be preferably the combination of cooling piece, radiator element.The further preferably combination of cooling piece, radiator element and exhaust fan.
In the present embodiment, heating piece 24 can directly be connected with sequencing reaction small chamber, is used for the sequencing reaction small chamber direct heating; Also can be connected with sequencing reaction small chamber through reflecting piece, the energy of heating piece 24 passes to sequencing reaction small chamber through reflecting piece, realizes the sequencing reaction small chamber heating.When sequencing reaction small chamber need heat; 24 pairs of sequencing reaction small chambers of heating piece heat, and temperature measuring equipment 21 is measured the temperature of sequencing reaction small chamber, and give DHE 20 with the Temperature Feedback of measuring; DHE produces steering order according to the temperature that receives; Make heating piece 24 carry out work, thereby realize heating sequencing reaction small chamber 1 according to steering order.Heating piece 24 continues sequencing reaction small chamber is heated, and up to reaching specified temperature, heating piece stops heating.Wherein, heating piece 24 is connected with DHE 20 through lead with temperature measuring equipment 21.Refrigeration plant 25 in the present embodiment is connected with liquid transmission assembly 4, is used for the reagent of liquid transmission assembly 4 is freezed, and can prolong the fresh keeping time of reagent, simultaneously, avoids user's frequent change reagent.Refrigerating reagent is transferred to sequencing reaction small chamber, can realize sequencing reaction small chamber is lowered the temperature.
Based on the foregoing description, the present invention proposes an embodiment, and is shown in figure 13.This gene sequencing equipment comprises liquid transmission assembly, sequencing reaction small chamber, temperature control assembly, light source assembly, adopts figure assembly and data handling component.Wherein:
(1) the liquid transmission assembly 4, are used for storage and transmission reagent.
(2) sequencing reaction small chamber 1, is used for fixing sample, and carries out sequencing reaction.
(3) the temperature control assembly 2, are fixed in the zone of sequencing reaction small chamber fixed sample, are used for sequencing reaction small chamber is carried out temperature control.
(4) light source assembly 3, are positioned at a sequencing reaction small chamber side relative with the temperature control assembly, are used to the light that provides photographic images required.
(5) adopt figure assembly 5, be used for photographic images.
(6) data handling component 6, are used to handle the image of adopting the shooting of figure assembly.
In the present embodiment, temperature control assembly 2 is positioned at a side of sequencing reaction small chamber 1, and light source assembly 3 is positioned at sequencing reaction small chamber 1 opposite side relative with the temperature control assembly.Sequencing reaction small chamber 1 can have a plurality of.Data handling component 6 can be connected the image of gathering in a plurality of figure of the adopting assemblies of PP simultaneously with a plurality of figure of adopting assemblies.Sequencing reaction small chamber 1 fixed sample of the gene sequencing equipment in the present technique scheme; Liquid transmission assembly 4 is transferred to sequencing reaction small chamber 1 with liquid, and 2 pairs of sequencing reaction small chambers of temperature control assembly 1 carry out temperature control, and sample and reagent carry out sequencing reaction in sequencing reaction small chamber 1; When reaction is accomplished; Light source assembly 3 provides photographic images required light, and light arrives sequencing reaction small chamber 2, adopts the image in the sequencing reaction small chamber after figure assembly 5 is taken sequencing reaction; Adopt figure assembly 5 image of taking is transferred to data handling component 6, data handling component 6 is handled and is adopted the image that figure assembly 5 is taken.
In the technical scheme of present embodiment, gene sequencing equipment has overcome the both sides that traditional shooting light field figure and the light source of taking fluorogram lay respectively at sequencing reaction small chamber 2, makes that the gene sequencing device structure is simple; The assembly 2 of temperature control simultaneously fully contacts with sequencing reaction small chamber 1, makes that temperature control efficient is higher; In addition, the zone that is used for fixing sample in the sequencing reaction small chamber is bigger, thereby makes that the flux of order-checking is higher.In addition, this gene sequencing equipment has been realized the robotization order-checking.
Need to prove; Gene sequencing equipment of the present invention adopts base complementrity paired principle to carry out nucleic acid sequencing; Each assembly of gene sequencing equipment in the foregoing description only is a preferred embodiment; All gene sequencing equipment based on same principle all is fit to and the present invention, also promptly all within protection scope of the present invention.
Should be noted that typical application of the present invention but be not limited to gene sequencing equipment itself, similarly in sequencing technologies, also can use device or the structure that the present invention sets forth at other.
The above is merely preferred embodiment of the present invention, not in order to restriction the present invention, all any modifications of within spirit of the present invention and principle, being done, is equal to and replaces and improvement etc., all should be included within protection scope of the present invention.
Claims (11)
1. a gene sequencing equipment is characterized in that, comprises sequencing reaction small chamber, temperature control assembly;
Said sequencing reaction small chamber is used for fixing sample, and carries out sequencing reaction;
Said temperature control assembly is fixed in the zone of sequencing reaction small chamber fixed sample, is used for sequencing reaction small chamber is carried out temperature control.
2. gene sequencing equipment according to claim 1 is characterized in that, said gene sequencing equipment also comprises light source assembly, is used to provide the light source of taking light field figure and the excitation light source of taking fluorogram;
Said light source assembly comprises light source, is positioned at a sequencing reaction small chamber side relative with the temperature control assembly, is used to the light that provides photographic images required.
3. gene sequencing equipment according to claim 2 is characterized in that said light source assembly also comprises reflecting piece and filtering apparatus;
Said reflecting piece between temperature control assembly and sequencing reaction small chamber, is used to reflect the light that arrives on the reflecting piece, takes the required light of light field figure to provide;
Said filtering apparatus is positioned at a side of light source, is used to filter stray light.
4. gene sequencing equipment according to claim 3 is characterized in that, said filtering apparatus also is used to change the light path through the light of said filtering apparatus.
5. gene sequencing equipment according to claim 3 is characterized in that the thermal conductivity of said reflecting piece is at least 80W/m.k, and its surface is at least 60% material by reflecting rate and processes.
6. gene sequencing equipment according to claim 1 is characterized in that, said gene sequencing equipment also comprises the liquid transmission assembly, is used for storage and transmission reagent.
7. gene sequencing equipment according to claim 6 is characterized in that, said liquid transmission assembly comprises device for storing liquid and passes liquid device;
Said device for storing liquid is used to store at least a reagent;
Said biography liquid device is positioned at a side of device for storing liquid, is used for the reagent of device for storing liquid is transferred to sequencing reaction small chamber.
8. according to each described gene sequencing equipment of claim 1 to 7, it is characterized in that said temperature control assembly comprises temperature measuring equipment, DHE, cooling piece and radiator element;
Said temperature measuring equipment is connected with sequencing reaction small chamber, is used for the temperature through the temperature sensor measurement sequencing reaction small chamber, and the temperature of measuring is sent to DHE;
Said DHE is connected with temperature measuring equipment, is used to receive and handle the temperature that temperature measuring equipment sends, and produces steering order;
Said cooling piece is connected respectively with sequencing reaction small chamber with DHE, is used for carrying out work according to steering order, so that the sequencing reaction small chamber heating or the required energy that freezes to be provided;
Said radiator element is connected with cooling piece, is used to transmit the energy of cooling piece one side.
9. according to claim 6 or 7 described gene sequencing equipment, it is characterized in that said temperature control assembly comprises temperature measuring equipment, DHE, heating piece and refrigeration plant;
Said temperature measuring equipment is connected with sequencing reaction small chamber, is used for the temperature through the temperature sensor measurement sequencing reaction small chamber, and the temperature of measuring is sent to DHE;
Said DHE is connected with temperature measuring equipment, is used to receive and handle the temperature that temperature measuring equipment sends, and produces steering order;
Said heating piece is connected respectively with DHE with sequencing reaction small chamber, is used for carrying out work according to steering order, to provide the sequencing reaction small chamber heating required energy;
Said refrigeration plant is positioned at liquid transmission assembly below, is used for the reagent in the liquid transmission assembly is freezed, and refrigerating reagent is transferred to sequencing reaction small chamber through the liquid transmission assembly, so that sequencing reaction small chamber is freezed.
10. according to each described gene sequencing equipment of claim 1 to 7, it is characterized in that the area that is used for fixing the zone of sample in the said sequencing reaction small chamber is S, 150mm
2≤S≤2100mm
2
11., it is characterized in that said sequencing reaction small chamber is hyperchannel or single passage sequencing reaction small chamber according to each described gene sequencing equipment of claim 1 to 7.
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