CN102321743B - Solid-phase carrier for surface fixation and preparation and application methods for solid-phase carrier - Google Patents
Solid-phase carrier for surface fixation and preparation and application methods for solid-phase carrier Download PDFInfo
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- CN102321743B CN102321743B CN201110222895.XA CN201110222895A CN102321743B CN 102321743 B CN102321743 B CN 102321743B CN 201110222895 A CN201110222895 A CN 201110222895A CN 102321743 B CN102321743 B CN 102321743B
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/16—Solid state fermenters, e.g. for koji production
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/16—Particles; Beads; Granular material; Encapsulation
Abstract
The invention relates to the field of surface chemical modification and genetic engineering and provides a solid-phase carrier for surface fixation and methods for preparing and applying the solid-phase carrier. The solid-phase carrier for surface fixation is connected with at least one surface group with an activation function, wherein the surface group contains a fixation end and an active end; and the molecular structural formula of the surface group is shown in the description: the fixation end is provided with at least one silyloxy group for combining the solid-phase carrier and the active end -R contains an active functional group for combining a surface fixation connection. In the invention, the solid-phase carrier is used for activation, and then resin is used for embedment and the solid-phase carrier is re-activated so as to ensure that the surface group on the surface of the solid-phase carrier is doubled, so that the quantity of the surface fixation connections fixed on the solid-phase carrier is increased.
Description
Technical field
The present invention relates to surface chemistry and genetically engineered field, more particularly, relate to a kind of for the fixing solid phase carrier in surface, and the preparation method of this solid phase carrier, and the application method of this solid phase carrier.
Background technology
DNA sequencing technology has been widely used in the every field of biological study, a lot of biological questions can be solved by high-throughput DNA sequencing technology, and extensive parallel order-checking platform (massively parallel DNA sequencing platform) has developed into the sequencing technologies of main flow.What at present, comparatively advanced sequenator used in the world is all new sequencing strategy---circulation chip sequencing (cyclic-array sequencing), also can be referred to as " new-generation sequencing technology or s-generation sequencing technologies ".
New-generation sequencing method is mostly by microemulsion PCR(emulsion PCR) or bridge-type PCR(bridge PCR) etc. method obtain parallel sequencing template.Microemulsion PCR, by water-in-oil System forming independent reaction space, need again to discharge recovery, and bridge-type PCR is relatively simple to operate, just solid phase carrier is had relatively high expectations after amplification.
The solid phase carrier that is applied to bridge-type PCR is that identical primer (nucleotide sequence) is fixed on to surface of solid phase carriers by certain reactive force, directly at surface of solid phase carriers, carries out PCR reaction.The solid phase carrier that is applied at present solid phase bridge-type PCR, its preparation method comprises the following steps: (1) utilizes the vitriol oil and hydrogen peroxide to make surface of solid phase carriers hydroxylation; (2) utilize aminopropyl triethoxysilane (APTES) to make surface silicon alkanisation; (3) utilize equal benzene nitrilotriacetic (BTA) to make surface of solid phase carriers activation; (4) by amino carboxyl effect immobilized oligonucleotide, pass through order-checking detection signal.Because fixed effect is not good, detecting during fluorescent signal, can only apply laser and excite to collect and enough signals detected.This testing conditions is too harsh, is unsuitable for widespread use.The problems such as above-mentioned solid phase carrier preparation method, exists complex operation, and fixed effect is poor, production cost height.
Therefore need a kind of new for fixing solid phase carrier in surface and preparation method thereof, improve quantity and the fixed efficiency of solid phase surface fixed group, the fast and convenient signal that detects surface and be fixedly connected with thing, can simplify procedures, reduce production costs simultaneously.
Summary of the invention
The object of the present invention is to provide and a kind ofly for fixing solid phase carrier in surface and its preparation method and application, be intended to solve solid phase surface fixed group on the low side, the problem that detection signal condition is too harsh, can simplify the operation simultaneously, reduces production costs.
In order to realize the object of the invention, the invention provides a kind ofly for the fixing solid phase carrier in surface, described solid phase carrier comprises that slide and at least one have the surface group of mobilizing function; Described surface group is directly connected with slide, comprises inboardend and active end, and its molecular structural formula is as follows:
Wherein, inboardend has at least one siloxy for being combined with slide.
Preferably, active end-R comprises for being fixedly connected with surface the activity functional groups that thing is combined, and wherein said activity functional groups is preferably combined with the other parts of active end-R by amido linkage; Described surface be fixedly connected with thing refer to by surface, be fixedly connected with after for detection of or detected material, include but not limited to oligonucleotide, albumen, antibody, probe.
Preferably, the activity functional groups that active end-R comprises has at least 2 for being fixedly connected with surface the end group that thing is combined.
Preferably, active end-R is the dendritic structure forming based on resin embedding, connects at least 2 for being fixedly connected with surface the end group that thing is combined.
Preferred, described is carboxyl for being fixedly connected with surface the end group that thing is combined.
Wherein, described surface of glass slide can be regular plane or the irregular surface with radian.
In order to realize the object of the invention, the present invention also provides a kind of preparation method for the fixing solid phase carrier in surface, and the described preparation method for the fixing solid phase carrier in surface comprises the following steps:
A. utilize silylating reagent to carry out end group modification to surface of glass slide;
B. utilize activating reagent in conjunction with end group, make activity functional groups on surface of glass slide band;
C. activity functional groups is carried out to resin embedding, form dendroid embedding body;
D. dendroid embedding body is activated again, obtain the solid phase carrier fixing for surface.
Wherein, described steps A comprises:
A1. slide is carried out to hydroxylation processing, make surface of glass slide carry hydroxyl;
A2. utilize silylating reagent and hydroxy combining, surface of glass slide is carried out to end group modification.
Preferably, surface of glass slide is carried out to the modification of amination end group.
Preferably, described step B is combined activating reagent by amido linkage with the amination end group of surface of glass slide.
Wherein, described activating reagent comprises activity functional groups.
Preferably, the activity functional groups that activating reagent comprises is C-terminal.
Preferably, when activity functional groups is C-terminal, described step C carries out embedding by polyamide resin to the C-terminal of surface of glass slide, forms dendritic structure.
Preferably, based on the activation treatment of described dendritic structure repeating step B, make slide connect at least 2 for being fixedly connected with surface the C-terminal that thing is combined.
In order to realize goal of the invention, the present invention also provides a kind of above-mentioned method of carrying out gene sequencing for the fixing solid phase carrier in surface of utilizing, and describedly utilizes above-mentioned method of carrying out gene sequencing for the fixing solid phase carrier in surface to comprise the following steps:
M1. by oligonucleotide fragment by being connected to solid phase carrier with the surface group effect with mobilizing function;
M2. the described solid phase carrier that is connected with oligonucleotide fragment is carried out to signal detection, by signal analysis, obtain base sequence information.
Wherein, the signal detection in described step M2 refers to the signal of measuring base to be measured by base complementrity principle, thereby obtains the sequence information of base to be measured.
Preferably, signal detecting method is including but not limited to connecting sequencing or synthetic sequencing.
In order to realize the object of the invention, the present invention also provides a kind of above-mentioned method of preparing gene chip for the fixing solid phase carrier in surface of utilizing, describedly utilize the above-mentioned method of preparing chip for the fixing solid phase carrier in surface to comprise: oligonucleotide fragment, by being connected to solid phase carrier with the surface group effect with mobilizing function, is formed to gene chip.
Wherein, the gene chip preparing described in can be used for the mensuration of DNA material capture and target nucleic acid sequence.
As from the foregoing, the present invention is by least 2 activity functional groups that comprise in the activity end-R connecting in surface of glass slide, add that resin embedding forms dendritic structure reactivate afterwards, reach the object that surface of glass slide activity functional groups is multiplied, thereby can fix multi-surface more and be fixedly connected with the object of thing, improve strength of signal, reduce the requirement to condition detection signal.Meanwhile, the above-mentioned method for the fixing solid phase carrier in surface of preparation of the present invention, operating process is simple, and production cost is low.
Accompanying drawing explanation
Fig. 1 is the structural representation for the surface group of the fixing solid phase carrier in surface in one embodiment of the invention;
Fig. 2 is the structural representation for the surface group of the fixing solid phase carrier in surface in one embodiment of the invention;
Fig. 3 is the structural representation for the surface group of the fixing solid phase carrier in surface in one embodiment of the invention;
Fig. 4 is the structural representation for the surface group of the fixing solid phase carrier in surface in one embodiment of the invention;
Fig. 5 carries out embedding structural representation afterwards for the surface group of the fixing solid phase carrier in surface in one embodiment of the invention;
Fig. 6 is the preparation method's schema for the fixing solid phase carrier in surface in one embodiment of the invention;
Fig. 7 is the preparation method's schema for the fixing solid phase carrier in surface in a preferred embodiment of the invention;
Fig. 8 is the method flow diagram based on carry out gene sequencing for the fixing solid phase carrier in surface in one embodiment of the invention;
Fig. 9 is based on carrying out for the fixing solid phase carrier in surface the order-checking fluorescent signal figure that gene sequencing obtains in one embodiment of the invention;
Figure 10 is based on carrying out method flow diagram prepared by gene chip for the fixing solid phase carrier in surface in one embodiment of the invention;
Figure 11 is the method flow schematic diagram based on carry out gene sequencing for the preparation of the fixing solid phase carrier in surface in one embodiment of the invention.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearer, below in conjunction with drawings and Examples, the present invention is further elaborated.
For the fixing solid phase carrier in surface, described solid phase carrier comprises that slide and at least one have the surface group of mobilizing function; Described surface group is directly connected with slide, comprises inboardend and active end, and its molecular structural formula is as follows:
Wherein, inboardend has at least one siloxy for being combined with slide.
Fig. 1 shows in the present invention for the universal architecture of the surface group of the fixing solid phase carrier in surface as follows:
Wherein, this surface group comprises inboardend and active end, and inboardend has at least one for be connected the siloxy using with slide; And active end-R comprises for being fixedly connected with surface the activity functional groups that thing is combined; This activity functional groups can be various forms, in following multiple embodiment, will elaborate the scheme that part is possible.
Fig. 2 shows in Fig. 1 the specific embodiment for the surface group of the fixing solid phase carrier in surface.This surface group structure is as follows:
In the present embodiment, the activity functional groups of the active end-R of surface group is connected with other parts of activity end-R by amido linkage.In addition, on phenyl ring, two remaining carboxyls become the activity functional groups in subsequent processing steps.
Fig. 3 shows in Fig. 1 the specific embodiment for the surface group of the solid phase carrier of surperficial immobilized oligonucleotide.This surface group structure is as follows:
In the present embodiment, activity functional groups part in the active end-R group of surface group is connected with other parts of active end-R by different sulphur cyanogen key, after different sulphur cyanogen key, it is the straight chain of three carbon atoms, with ester bond, be connected with ring texture, in ring texture, remaining carboxyl is as the activity functional groups of follow-up use.
Fig. 4 shows in Fig. 1 the specific embodiment for the surface group of the solid phase carrier of surperficial immobilized oligonucleotide.This surface group structure is as follows:
In the present embodiment, surface group inboardend is connected by two siloxies with active end-R, activity functional groups part in active end-R is connected with other parts of active end-R by amido linkage, encircles the activity functional groups of remaining two carboxyls as follow-up use.
It should be noted that Fig. 2, Fig. 3 and Fig. 4 have only provided several specific embodiments of surface group, but the present invention is not so limited.In principle, with the two or more surface groups that can be fixedly connected with surface the activity functional groups that thing is connected.
Fig. 5 shows in an embodiment for the structure after the universal architecture process subsequent disposal of the fixing solid phase carrier in surface.Activity end-R after preferably surface of glass slide being activated by polyamide resin carries out embedding, makes the surface group of surface of glass slide combination form dendritic structure.Further, after activation, on dendritic structure, reactivate into the activity functional groups of original active end-R, thereby make the increase of its quantity generation multiple, can fix more surface and be fixedly connected with thing.
Fig. 6 has provided the method flow of preparing for the fixing solid phase carrier in surface in one embodiment of the invention.Present method comprises the following steps:
S1. utilize silylating reagent to carry out end group modification to surface of glass slide;
S2. utilize activating reagent to be combined with end group, surface of glass slide is connected for being fixedly connected with surface the activity functional groups that thing is combined;
S3. activity functional groups is carried out to resin embedding, form dendroid embedding body;
S4. dendroid embedding body is activated again, obtain the solid phase carrier fixing for surface.
It should be noted that, before starting in steps, slide used is cleaned and removes impurity, then surface of glass slide is carried out to hydroxylation processing, make hydroxyl on surface of glass slide band.Described surface of glass slide can be regular plane or irregular as with the surface of radian.In one embodiment of the invention, the glass of alternation rule plane is as slide.
Wherein, hydroxylation is processed the processing of preferred alcohols alkaline process, also can select simultaneously other as methods such as sour oxygen methods (vitriol oil and dioxygen water law).About hydroxylated preferred version, will be described in detail in another preferred embodiment.
In step S1, surface of glass slide is carried out to end group modification.Utilize alkylating reagent to react with the hydroxyl on slide, form amidized siloxy inboardend.
Wherein, silylating reagent in step S1, can select in principle siloxanes or siloxanes acid, dioxane (dioxane) solution that therefore can choice for use carbonyl dimidazoles (CDI), 3-Racemic glycidol propyl trimethoxy silicane (GOPS) the toluene liquid that contains triethylamine (TEA), aminopropyl triethoxysilane (APTES) are as the reagent of silanization.For the preferred version of step S1, can in follow-up preferred embodiment, provide detailed description.
In step S2, utilize activating reagent, make activity functional groups on surface of glass slide band.Surface of glass slide siloxy inboardend is amination when end modified, preferably by amido linkage, activating reagent is combined with the amination end group of surface of glass slide, thereby makes surface of glass slide band upper surface group-R.
Wherein, active end-R can be fixedly connected with the activity functional groups that thing is connected with surface with two or more.Therefore, can select different activating reagents, for example can select saturated Pyroglutaric acid (DMF)/NHS/N, N'-dicyclohexyl carbodiimide (DCC), 3-Racemic glycidol propyl trimethoxy silicane/silane coupling agent (GOPTS), inferior phenylene diisothiocyanic acid salts solution (PDITC) are as activating reagent.About the preferred version of step S2, will in follow-up preferred embodiment, provide.
In step S3, the activity end-R of effects on surface group carries out resin embedding, forms dendroid embedding body.By resin embedding, the activity functional groups that activity end-R is comprised carries out numerical value amplification, in the dendritic structure that simultaneously guarantees to form, connects at least 2 for being fixedly connected with surface the activity functional groups that thing is combined.
In step S4, the operation that dendroid embedding body is activated repeats according to the method for step S2.
Above-mentioned for the fixing solid phase carrier in surface in the present invention, its surface be fixedly connected with thing refer to by surface of glass slide, fixed after for detection of or detected material, include but not limited to oligonucleotide, albumen, antibody and probe.
Fig. 7 has provided the method flow of a preferred embodiment of preparing for the fixing solid phase carrier in surface of the present invention.Concrete step comprises:
S101. adopt NaOH and ethanol to process, make surface of glass slide hydroxylation;
S102. utilize silylating reagent APTMS, make the hydroxyl amino of surface of glass slide;
S103. utilize activator TMA/NHS/DCC to activate, make activity functional groups on surface of glass slide band;
S104. with polymeric amide PAMAM to activity functional groups embedding, form dendroid embedding body;
S105. dendroid embedding body is activated again, obtain the solid phase carrier fixing for surface.
It should be noted that, before starting in steps, need to clean testing slide used, remove the impurity of surface of glass slide.
In step S101, preferably by alcohol alkaline process, make hydroxyl on surface of glass slide band.Specific operation process is: the NaOH solution of preparation 18%, then joins mix and blend in ethanol solution until liquid is limpid; Slide is put into wherein, isolated air, 2h is soaked in vibration, then takes out slide washed several times with water and removes remaining alcohol, alkali, dries up.
In this step, the concrete reaction formula relating to is as follows:
In step S102, utilize silylating reagent to process slide and make its surface hydroxyl amination.Preferably the acetone solution of 1% aminopropyl trimethoxysilane (APTMS) carries out Silanization reaction.Concrete operations are: slide is put into 1% aminopropyl trimethoxysilane (APTMS) and 95% acetone solution and soak 2min, make on its surface band-NH
2, then acetone oscillation cleaning, N
2dry.Concrete chemical reaction is as follows:
In step S103, utilize activating reagent to process surface of glass slide and make its surface active and be with activity functional groups.Preferably trimesic acid (TMA), N-hydroxy-succinamide (NHS), N, N '-dicyclohexylcarbodiimide (DCC) solution and diisopropylethylamine (DIEA), as surfactant, make surface of glass slide activation, are with activity functional groups.
Wherein, concrete operations comprise: with trimesic acid (TMA), N-hydroxy-succinamide (NHS), N, air-isolation oscillatory reaction 2h after N '-dicyclohexylcarbodiimide (DCC) solution mixes, centrifugation product, is mixed with into Acibenzolar with diisopropylethylamine (DIEA) solution equal-volume; Then at the slide one side of silanization, drip appropriate Acibenzolar, reaction 3h makes surface of glass slide radical functino, N, N-dimethylformamide (DMF) and ethanol oscillation cleaning.Then use the sodium hydrogen carbonate solution of 5%, pH8.8 to process, make carboxyl on surface of glass slide band.Concrete chemical equation is as follows:
In step S104, by polymeric amide PAMAM resin, the carboxyl of surface of glass slide is carried out to embedding, form dendroid embedding body.Drip methanol solution that 100 μ l are dissolved with 10% polymeric amide PAMAM in surface of glass slide, then be combined formation sandwich structure with second slide, room temperature is put 12h.Then use washed with methanol 2 times, N2 is dry.
The related reaction of step S104 is as follows:
In step S105, the operation that dendroid embedding body is activated repeats according to the method for step S103.
Fig. 8 has provided solid phase carrier based on a kind of surperficial immobilized oligonucleotide in one embodiment of the invention and has been applied to the method flow of gene sequencing.Concrete step comprises:
In step S201, carry out the point sample of testing sample.Testing sample mixes with order-checking buffer, carries out point sample on the solid phase carrier for preparing and be positioned in container.
Wherein, when biased sample, can add a small amount of microballon, for focusing on, make sample can concentrate on certain position place.
In step S202, sealing lucifuge is carried out the room temperature of testing sample and is fixed.Utilize masking foil to encase lucifuge by being placed with the container of putting excellent solid phase carrier, under room temperature, fix 16h.
In step S203, loading is carried out gene sequencing, detection signal.Solid phase carrier after fixing end, washs with TE, and uses ddH
2o washed.Washed solid phase carrier is packed in order-checking cell, carry out gene sequencing, collect fluorescent signal.
Wherein, gene order surveying method described in step S103, can be to connect sequencing, can be also synthetic sequencing.Much more no longer the two concrete principle and operation about this, can, with reference to existing detailed description in prior art, repeat at this.
Wherein, the fluorescent signal result figure obtaining according to connection sequencing as shown in Figure 9.
Figure 10 has provided the method flow that carries out gene chip Application and preparation in one embodiment of the invention based on a kind of solid phase carrier of surperficial immobilized oligonucleotide.Concrete step comprises:
In step S301, by being fixed on solid phase carrier by the mobilizing function group of surface of glass slide with the oligonucleotide fragment of target DNA complementation, be prepared into gene chip.
In step S302, testing sample and gene chip are hatched to combination, catch target DNA fragmentation.
In step S303, after catching end, reaction system is washed with damping fluid, remove the testing sample not being attached on gene chip, thereby obtain target DNA fragmentation.
Figure 11 is based on the preparation of a kind of solid phase carrier for immobilized oligonucleotide and be applied to the schematic diagram of gene order surveying method flow process in one embodiment of the invention.This figure has represented the preparation process based on a kind of solid phase carrier for immobilized oligonucleotide intuitively, and then carries out the fixing also process of detection signal of oligonucleotide:
The mixed solution of step 3, employing activating reagent TMA/NHS/DIEA, carries out condensation reaction by the amino of its carboxyl and surface of glass slide, makes surface of glass slide form activity functional groups; And then pass through NaHCO
3processing, make its activity functional groups become pendant carboxylic group.
Step 4, take polymeric amide PAMAM resin, carboxyl (in the present embodiment-R group as carboxyl) is carried out to embedding, and then to resin surface with amino according to the method in step 3, reactivate, make the pendant carboxylic group quantity of surface of glass slide become multiple to increase.
The condensation of step 5, utilization amino and carboxyl, is connected and fixed oligonucleotide fragment in the surface of glass slide of aforesaid method activation, then by connection, checks order or synthesizes the method checking order and carry out the detection of signal.
It should be noted that the typical application of the present invention but be not limited to the fixing amplification of oligonucleotide, at other similar protein molecules and other, containing in amino compound, also can apply method set forth in the present invention.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any modifications of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.
Claims (11)
1. for the fixing solid phase carrier in surface, it is characterized in that, described solid phase carrier comprises that slide and at least one have the surface group of mobilizing function; Described surface group is directly connected with slide, comprises inboardend and active end, and its molecular structural formula is as follows:
Described inboardend has at least one siloxy for being combined with slide;
Described active end-R comprises for being fixedly connected with surface the activity functional groups that thing is combined.
2. according to claim 1ly for the fixing solid phase carrier in surface, it is characterized in that, described activity functional groups is combined with the other parts of active end-R by amido linkage.
3. according to claim 1 and 2ly for the fixing solid phase carrier in surface, it is characterized in that, described activity functional groups has at least 2 for being fixedly connected with surface the end group that thing is combined.
4. according to claim 3ly for the fixing solid phase carrier in surface, it is characterized in that, described active end-R is the dendritic structure based on resin embedding formation, connects at least 2 for being fixedly connected with surface the end group that thing is combined.
5. according to claim 3ly for the fixing solid phase carrier in surface, it is characterized in that, described surface of glass slide is regular plane or the irregular surface with radian.
6. a preparation method for solid phase carrier as claimed in claim 1, is characterized in that, said method comprising the steps of:
A. utilize silylating reagent to carry out end group modification to surface of glass slide;
B. utilize activating reagent in conjunction with end group, make activity functional groups on surface of glass slide band;
C. activity functional groups is carried out to resin embedding, form dendroid embedding body;
D. dendroid embedding body is activated again, obtain the solid phase carrier fixing for surface;
Described steps A comprises:
A1. by alcohol alkaline process, slide is carried out to hydroxylation processing, make surface of glass slide carry hydroxyl;
A2. utilize silylating reagent and hydroxy combining, surface of glass slide is carried out to end group modification.
7. the preparation method of solid phase carrier according to claim 6, is characterized in that, if surface of glass slide is carried out to amidized end group modification in steps A 2, described step B comprises:
By amido linkage, activating reagent is combined with the amination end group of surface of glass slide;
Wherein, described activating reagent comprises activity functional groups.
8. the preparation method of solid phase carrier according to claim 7, is characterized in that, if the activity functional groups that activating reagent comprises is C-terminal, step C comprises:
C1. by polyamide resin, the C-terminal of surface of glass slide is carried out to embedding, form dendritic structure;
C2. the activation treatment based on described dendritic structure repeating step B, makes slide connect at least 2 for being fixedly connected with surface the C-terminal that thing is combined.
9. utilize solid phase carrier described in claim 1 to carry out a method for gene sequencing, it is characterized in that, described method comprises:
M1. by oligonucleotide fragment by being connected to solid phase carrier with the surface group effect with mobilizing function;
M2. the described solid phase carrier that is connected with oligonucleotide fragment is carried out to signal detection, and by the signal in testing process is analyzed, thereby base sequence information obtained.
10. the method for gene sequencing according to claim 9, is characterized in that, the signal detection in described step M2 refers to the signal of measuring base to be measured by base complementrity principle, thereby obtains the sequence information of base to be measured.
11. 1 kinds of methods of utilizing solid phase carrier described in claim 1 to prepare gene chip, is characterized in that, described method is: oligonucleotide fragment, by being connected to solid phase carrier with the surface group effect with mobilizing function, is formed to gene chip.
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