CN106198952A - A kind of suppress the nucleic acid molecules enclosure method to sensing interface non-specific adsorption - Google Patents
A kind of suppress the nucleic acid molecules enclosure method to sensing interface non-specific adsorption Download PDFInfo
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- CN106198952A CN106198952A CN201610509843.3A CN201610509843A CN106198952A CN 106198952 A CN106198952 A CN 106198952A CN 201610509843 A CN201610509843 A CN 201610509843A CN 106198952 A CN106198952 A CN 106198952A
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- salmon sperm
- sperm dna
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/5306—Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
Abstract
The invention discloses and a kind of suppress the nucleic acid molecules enclosure method to sensing interface non-specific adsorption.Enclosure method disclosed by the invention includes: process sensor with salmon sperm dna, it is achieved the closing of sensor；Sensor is processed with salmon sperm dna solution；The solvent of salmon sperm dna solution is PBS, and solute is salmon sperm dna, and in salmon sperm dna solution, the concentration of salmon sperm dna is 0.01mg/mL 10mg/mL；PBS is made up of solvent and solute, solvent be water, solute and concentration thereof be 0.1M disodium hydrogen phosphate and sodium dihydrogen phosphate and 0.15M sodium chloride, pH is 7.4.It is demonstrated experimentally that the signal ratio using SSDNA to close the specific binding and non-specific adsorption of nucleic acid molecules on the sensing interface of strategy is respectively without closing, traditional BSA closes and 40.20,6.93 and 4.23 times of simple A oligomer closing strategy.
Description
Technical field
The present invention relates to a kind of in biological technical field suppress the nucleic acid molecules closing to sensing interface non-specific adsorption
Method.
Background technology
In classical immune analysis method, the affine identification between antigen-antibody constitutes the basis of sensing.Affine work
With being a series of noncovalent forces (hydrophobe effect, electrostatic interaction, hydrogen bond etc.) between part and receptor under specific biological molecules structure
General name, the comprehensive function of multiple low force ensure that the stable bond of affine both sides.But, non-mesh present in the complex system
Mark material may produce absorption to affinity ligand and receptor to varying degrees, is referred to as " non-specific adsorption ".At actual experiment
During operation designs with related device, generally need to add the avtive spot knot of sealer (blocking agent) and sensing interface
Close, to reduce non-specific adsorption, simultaneously the most not being normally carried out of target compatible reaction in interference system.Affine instead at traditional immunization
Ying Zhong, is generally served as sealer by protein cheap and easy to get, such as bovine serum albumin (BSA) and ovalbumin (OVA) etc..
Outside the compatible reaction that removing protein participates in, nearly more than two decades comes, using functional nucleic acid as new bio recognition component
Sensing analytical method also achieves swift and violent development.Functional nucleic acid is the oligomerization core with specific functions such as affine identification, catalysis
Acid fragments, with aptamer (aptamer) with two kinds of nucleic acid material of DNA enzymatic (DNAzyme) as representative.The most existing over one hundred
Planting functional nucleic acid to be in the news, it is up to a hundred that it identifies that object contains metal ion, biotoxin, cell metabolite, high molecular weight protein etc.
Planting material, its application concentrates on the aspect such as bio-sensing and living cells diagnosis and treatment.
When building sensing interface based on functional nucleic acid, need also exist for " the non-spy evading nucleic acid molecules to sensing interface
Opposite sex absorption ", carry out sealing treatment for different sensing systems.In immune sensing, widely used protein blocking material exists
During locked nucleic acids interface not enough: 1) participate in important reaction or as in the system of target at metal ion, protein blocking
The existence of layer may interfere with the carrying out of sensing process；2) albumen is the polymer macromolecule formed by amino acid condensation, table
Surface charge distribution is uneven, there may be stronger local adsorption site (such as basic amine group for flexible, thread nucleic acid material
The acid Electrostatic Absorption to elecrtonegativity nucleic acid chains).Therefore, protein material the optimal choosing of non-inhibited nucleic acid molecules non-specific adsorption
Select.
2016, the Juewen Liu seminar at Canada research in nanotechnology center, Waterloo was a kind of based on homogeneous phase solution
Graphene-nucleic acid complexes sensing system is systematically probed into ten kinds of surface-closed materials to nucleic acid molecules non-specific adsorption
Inhibition；Main closed material involved by this seminar includes that high molecular polymer, surfactant and nucleic acid are short
Chain is (with A15For representing), its research shows, electrostatic interaction plays important work during the non-specific adsorption of suppression nucleic acid
With；Additionally, nucleic acid short chain is the potential closed material of a kind of tool, nucleic acid chains length is the longest, is combined with the closing of sensing surface
The strongest.But, the research system of this seminar is immobilized in graphenic surface by pi-pi accumulation effect absorption by pickup probe nucleic acid,
There is the probability of pickup probe nucleic acid and locked nucleic acids competition graphenic surface site；Additionally, Graphene-nucleic acid system is at body
Sensing in phase solution, device is difficult to reuse.It is true that design compared to the pickup probe in homogeneous phase solution, based on
The biosensor of the interfacial structures such as optical fiber, chip of light waveguide, electrode has immobilized stable, the integrated height of probe, interface can be anti-
The advantages such as multiple utilization (single testing cost is low), monitoring can be carried out in real time, meet the requirement to biosensor on stricti jurise.
Wherein, probe is immobilized stably refers to based on sensing interface by covalency or the immobilized probe of affine mode, than adsorbing immobilized strategy
The most strong durable.In existing nucleic acid sensor based on interface, non-specific adsorption situation the most generally exists.At present
There is no pervasive high-quality sealer and be seen in report.
Summary of the invention
The technical problem to be solved is how to suppress nucleic acid molecules to sensing interface non-specific adsorption, comes real
The closing of existing sensing interface.
For solving above-mentioned technical problem, present invention firstly provides a kind of suppression nucleic acid molecules non-specific to sensor interface
Property absorption enclosure method.
The suppression nucleic acid molecules provided by the present invention enclosure method to sensor interface non-specific adsorption, including using salmon
Milt DNA (SSDNA) processes sensor, it is achieved the closing of sensor.
In above-mentioned enclosure method, described salmon sperm dna is the salmon sperm dna of degeneration, such as the salmon of sonicated degeneration
Essence DNA.Described salmon sperm dna can use phenol chloroform to obtain.Described salmon sperm dna concretely Suo Laibao (solarbio)
The SSDNA that article No. is H1060.The salmon sperm dna of described degeneration refers to long chain interruption, forms the salmon essence of many short pieces
DNA。
In above-mentioned enclosure method, described salmon sperm dna processes sensor and can include with the process of salmon sperm dna solution described
Sensor；Solute in described salmon sperm dna solution is salmon sperm dna.Described salmon sperm dna solution is by described solute and solvent
Composition, as long as described solvent meets described closing and requires.
In above-mentioned enclosure method, the solvent of described salmon sperm dna solution can be PBS；Described PBS is by molten
Agent and solute composition, described solvent is water, and described solute and concentration thereof are 0.1M disodium hydrogen phosphate and sodium dihydrogen phosphate, 0.15M
Sodium chloride, pH is 7.4.Wherein 0.1M disodium hydrogen phosphate and sodium dihydrogen phosphate represent disodium hydrogen phosphate and phosphorus in PBS
The total concentration of acid dihydride sodium is 0.1M.
In above-mentioned enclosure method, described salmon sperm dna concentration in described salmon sperm dna solution is 0.01mg/mL-
10mg/mL.Described salmon sperm dna concentration concretely 0.01mg/mL-0.1mg/mL in described salmon sperm dna solution, as
0.05mg/mL。
In above-mentioned enclosure method, described salmon sperm dna processes sensor or described salmon sperm dna solution processes described
The time of sensor can be all 2-5 hour, such as 2 hours.
In above-mentioned enclosure method, described salmon sperm dna processes sensor or described salmon sperm dna solution processes described
Sensor all can be carried out at 30-37 DEG C.
In above-mentioned enclosure method, described sensor can be biosensor or Fibre Optical Sensor.Described biosensor is
Referring to utilize biomaterial to make the sensor of recognition component, common biomaterial includes: antibody, functional protein, full cell, nucleic acid
Deng.
In above-mentioned enclosure method, the most following a1 of described sensor) or a2) or a3) or a4):
A1) evanescent wave optical fiber sensor；
A2) nucleic acid sensor；
A3) optical fiber biosensor；
A4) evanescent wave optical fiber biosensor.
For solving above-mentioned technical problem, present invention also offers salmon sperm dna non-to sensor interface at suppression nucleic acid molecules
Application in specific adsorption.
In above-mentioned application, described sensor can be biosensor or Fibre Optical Sensor.Described biosensor refers to profit
Make the sensor of recognition component of biomaterial, common biomaterial includes: antibody, functional protein, full cell, nucleic acid etc..
In above-mentioned application, the most following a1 of described sensor) or a2) or a3) or a4):
A1) evanescent wave optical fiber sensor；
A2) nucleic acid sensor；
A3) optical fiber biosensor；
A4) evanescent wave optical fiber biosensor.
In the present invention, the optical fiber of described optical fiber concretely 600 μMs of core diameters.The optical fiber of these 600 μMs of core diameters is the most southern
Jing Chun brightness Scientific and Technical Industry Co., Ltd product.
Sensing interface after modifying, with salmon sperm dna as sealer, is closed under the conditions of 37 DEG C by the present invention.Close
After interface can keep stable in the presence of flowing mutually, and effectively inhibit the non-specific suction of non-targeted nucleic acid
Attached.Specifically (Fig. 4), in the case of not closing, sensing surface presents more avtive spot, it is easy to combine in liquid phase
Non-targeted nucleic acid, forms the strongest non-specific adsorption；In the system using salmon sperm dna to close, not only most alive
Property site is closed by salmon sperm dna, electrostatic repulsion (salmon sperm dna and the liquid phase middle reaches between more Presence of an interface and free nucleic acid
Freestone acid all presents elecrtonegativity, and the same sex is repelled each other), therefore the non-specific adsorption of sensing interface is effectively suppressed by nucleic acid.Use
SSDNA closes the signal of the specific binding and non-specific adsorption of nucleic acid molecules ratio on the sensing interface of strategy and is respectively without envelope
Close, traditional BSA closes and simple A oligomer closes tactful 40.20,6.93 and 4.23 times, uses SSDNA to close plan
The effect slightly closed sensing interface is significantly better than without closing, traditional BSA closes and simple A oligomer closes plan
Slightly；And use SSDNA to carry out closing and there is economy, simplicity, quick advantage.The present invention is applicable to all depend on solid-liquid circle
Face (such as a kind of solid liquid interface of optical fiber-liquid to be measured in the present invention) carries out the biosensor detected.
Accompanying drawing explanation
Fig. 1 is the method that the present invention contrasts specific adsorption and non-specific adsorption signal.
Fig. 2 is the course of reaction of synthesis BSA-DTB complex.
Fig. 3 is the course of reaction that BSA-DTB complex modifies optical fiber.
Fig. 4 is that SSDNA of the present invention suppresses the nucleic acid molecules schematic diagram to the enclosure method of sensing interface non-specific adsorption.
Fig. 5 is the result that the present invention reduces non-specific adsorption effect.
Detailed description of the invention
Being further described in detail the present invention below in conjunction with detailed description of the invention, the embodiment be given is only for explaining
The bright present invention rather than in order to limit the scope of the present invention.
Experimental technique in following embodiment, if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, the most commercially obtain.
PBS in following embodiment is made up of solvent and solute, and solvent is that water, solute and concentration thereof are
0.1M0.1M disodium hydrogen phosphate and sodium dihydrogen phosphate (i.e. the two total concentration in PBS is 0.1M), 0.15M chlorination
Sodium, pH 7.4.
Eluent in following embodiment is SDS aqueous solution, and wherein the mass percent of SDS is 0.05%, adjusts with hydrochloric acid
Whole pH is 1.9；SDS is amresco product, and article No. is M107.
Embodiment 1, utilize SSDNA that sensing interface is closed
The important indicator evaluating non-specific adsorption degree is the specific binding signal ratio with non-specific adsorption.
The present embodiment utilizes the parent of Streptavidin (Streptavidin, STV) and desthiobiotin (Desthiobiotin, DTB)
Right with the identification specific binding identification of structure.As it is shown in figure 1, first build the optical fiber that desthiobiotin is modified, this optical fiber is permissible
The specific binding nucleic acid conjugate probe (STV-DNA-Cy5.5) containing Streptavidin, obtains specific binding signal；With
Time, utilize free Streptavidin that matched group is set with free nucleic acid, investigate non-specific binding signal.Contrast specificity knot
Close the ratio of signal and non-specific binding signal, the non-specific adsorption degree of system can be evaluated.
One, synthesis BSA-DTB complex
In view of the characteristic of evanescent wave optical fiber system, build structure sheaf with shielding flowing middle fluorescence molecule mutually at optical fiber surface
The fluorescence interference being likely to result in, the present invention is with economical and easily available bovine serum albumin as structural material, as the load of desthiobiotin
Body, prepares bovine serum albumin-desthiobiotin complex (BSA-DTB complex).By a series of cross-linking agent, BSA-DTB is multiple
Compound is modified at optical fiber surface.Wherein, the method for BSA-DTB complex is synthesized as in figure 2 it is shown, BSA-DTB represents BSA-in Fig. 2
DTB complex: utilize BSA surface amino groups and desthiobiotin derivant NHS desthiobiotin (NHS-DTB, Thermo,
Article No.: 16129) carry out conjugation reaction.Concrete preparation method is as follows:
In PBS add BSA obtain the BSA solution that BSA concentration is 4mg/mL, in BSA solution add put to
The NHS-DTB of room temperature, the mol ratio making NHS-DTB Yu BSA is 10:1, reacts 1h at room temperature；Reacted mixed liquor is (main
BSA-DTB target product to be comprised and the little molecule of unreacted NHS-DTB) add in batches in Amicon-10K super filter tube,
Carry out initial centrifugation purification (12000rpm, centrifugal 10min)；After preliminary purification, then in inner tube, add 200 μ L PBS bufferings
Liquid, is centrifuged 10min in 12000rpm, collects inner tube liquid, it is thus achieved that the BSA-DTB complex of purification, in 4 DEG C of guarantors after being repeated 8 times
Deposit.
Two, the optical fiber that desthiobiotin is modified is made
With Piraha solution (by dense H2SO4With commercially available 30% (mass percent concentration) H2O2Composition, dense H2SO4With
30%H2O2Volume ratio be 3:1, wherein, dense H2SO4The concentrated sulphuric acid of 98% produced for Beijing Chemical Plant) soak at 120 DEG C
Optical fibers (be called for short optical fiber, 600 μMs of core diameters (Nanjing Chunhui Science and Technology Industrial Co Ltd)), this optical fiber in an experiment be passed through sharp
Under conditions of light, surface can form evanscent field) one hour, then fully wash optical fiber neutral (pH is about to washing liquid with ultra-pure water
7), and dry up optical fiber with nitrogen, be put in 70 DEG C of baking ovens overnight；The body of secondary daily 3-mercaptopropyi trimethoxy silane (MTS)
Long-pending percent concentration be 2% MTS solution (solvent of this MTS solution is dry toluene) at room temperature soak optical fiber two hours,
Clean for several times with dry toluene afterwards；Then 4-maleimidobutyric acid-N-succinimide ester (GMBS) is placed the fiber in
The GMBS solution that concentration is 1mg/mL (solvent of this GMBS solution is ethanol) in room temperature reaction 1h, reacted optical fiber is used
Alcohol flushing three times, then rinse well with high purity water, obtain stand-by optical fiber.
Utilizing ultraviolet spectrophotometer to carrying out quantitatively, BSA-DTB complex step one obtained adds to PBS
In, obtain the BSA-DTB solution (concentration of this BSA-DTB solution is in terms of albumen (i.e. BSA) comes) that concentration is 0.1mg/mL；
Stand-by optical fiber is put in BSA-DTB solution, it is ensured that the BSA-DTB in BSA-DTB solution is excessive relative to optical fiber, in 4
DEG C reaction overnight (12 hours).After cleaning reacted optical fiber with deionized water next day, in 4 DEG C of preservations, (overall modification is as schemed
Shown in 3), obtain the optical fiber that BSA-DTB modifies.
Three, detection SSDNA sealing effect
One) preparation of SSDNA:
Being dissolved by SSDNA (solarbio product, article No. is H1060) PBS, obtaining SSDNA concentration is
The SSDNA solution of 0.05mg/mL.Wherein, the article No. of Solarbio is that the SSDNA of H1060 is for utilize phenol chlorine from salmon sperm
Imitative extracting the salmon sperm dna of sonicated degeneration, the long chain interruption of salmon sperm dna of this degeneration, form many short pieces.
Two) SSDNA closes optical fiber
This step is selected biotin-A20-Cy5.5 (BA20) is experimental group target nucleic acid, with A20-Cy5.5 (A20) is
Blank group nucleic acid, experiment is in triplicate.Wherein, BA20 5 ' end to 3 ' end sequences as shown in sequence 1 in sequence table,
5 ' ends of BA20 sequence are by biotin labeling, and 3 ' ends are by Cy5.5 labelling；5 ' the ends of A20 are to sequence in the sequence such as sequence table of 3 ' ends
Shown in row 1,3 ' ends of BA20 sequence are by Cy5.5 labelling.
The optical fiber that BSA-DTB step 2 obtained modifies is divided into 5 groups, is respectively designated as matched group, None by these five groups
Group, BSA group, A20 group and SSDNA group.
1, SSDNA group is processed as follows:
Close the optical fiber two hours of the BSA-DTB modification of SSDNA group at 37 DEG C with the SSDNA solution of 0.05mg/mL, obtain
The optical fiber that SSDNA closes.
By biotin-A20-Cy5.5 (BA20) at room temperature buffers in PBS with the Streptavidin (STV) with molar concentration
Liquid reacts 20 minutes, obtains reactant liquor BA20-STV, make the biotin (biotin) on BA20, with STV, affine identification occur,
STV still possesses unnecessary affine site and can combine with the desthiobiotin molecule of optical fiber surface simultaneously.By A20 and same molar concentration
STV at room temperature in PBS react 20 minutes, obtain reactant liquor A20-STV.
Reactant liquor BA20-STV is passed through evanescent wave optical fiber sensing platform, and (evanescent wave optical fiber sensing platform is Chinese patent Shen
Please number be the biosensor of full fiber optic evanescent wave of 200610089497.4).During fiber laser arrays, first buffer with PBS
Liquid rinses the optical fiber of SSDNA closing until baseline is steady；After baseline is steady, it is liquid to be measured by reactant liquor BA20-STV, opens compacted
Dynamic pump sample introduction 15s；Stop peristaltic pump afterwards, make liquid to be measured react 90s in reaction tank；It is again turned on peristaltic pump, is passed through eluting
Liquid；Finally it is passed through PBS, steady to baseline, stop sampling simultaneously, (these data are specific binding signal to preserve data
With the mixed signal of non-specific signals, by its named Sa+b/SSDNA)。
According to the method described above, BA20-STV being replaced with A20-STV, as comparison, other steps are the most constant, obtain non-spy
Specific signal, by its named Sb/SSDNA.Specific binding signal and the ratio=(S of non-specific signalsa+b/SSDNA-Sb/
SSDNA)÷Sb/SSDNA。
2, A20 group is processed as follows:
Being dissolved by A oligomer (sequence is as shown in sequence 1 in sequence table) PBS, obtaining A oligo concentrations is
The A oligo solution of 0.05mg/mL.Close the BSA-DTB modification of A20 group with the A oligo solution of 0.05mg/mL at 37 DEG C
Optical fiber two hours, obtains the optical fiber that A oligomer is closed.
The reactant liquor BA20-STV of step 1 is passed through evanescent wave optical fiber sensing platform.During fiber laser arrays, first use
PBS rinses the optical fiber of A oligomer closing until baseline is steady；After baseline is steady, it is to be measured by reactant liquor BA20-STV
Liquid, opens peristaltic pump sample introduction 15s；Stop peristaltic pump afterwards, make liquid to be measured react 90s in reaction tank；It is again turned on peristaltic pump,
It is passed through eluent；Finally it is passed through PBS, steady to baseline, stop sampling simultaneously, (these data are specificity to preserve data
Binding signal and the mixed signal of non-specific signals, by its named Sa+b/A20)。
According to the method described above, BA20-STV replacing with the A20-STV of step 1, as comparison, other steps are the most constant,
Obtain non-specific signals, by its named Sb/A20.Specific binding signal and the ratio=(S of non-specific signalsa+b/
A20-Sb/A20)÷Sb/A20。
3, BSA group is processed as follows:
BSA PBS is dissolved, obtains the BSA solution that BSA concentration is 0.05mg/mL.At 37 DEG C with 0.05mg/
The optical fiber that the BSA-DTB of the BSA solution closing BSA group of mL modifies two hours, obtains the optical fiber that BSA closes.
The reactant liquor BA20-STV of step 1 is passed through evanescent wave optical fiber sensing platform.During fiber laser arrays, first use
PBS rinses the optical fiber of BSA closing until baseline is steady；After baseline is steady, it is liquid to be measured by reactant liquor BA20-STV, opens
Open peristaltic pump sample introduction 15s；Stop peristaltic pump afterwards, make liquid to be measured react 90s in reaction tank；It is again turned on peristaltic pump, is passed through
Eluent；Finally it is passed through PBS, steady to baseline, stop sampling simultaneously, (these data are specific binding to preserve data
Signal and the mixed signal of non-specific signals, by its named Sa+b/BSA)。
According to the method described above, BA20-STV replacing with the A20-STV of step 1, as comparison, other steps are the most constant,
Obtain non-specific signals, by its named Sb/BSA.Specific binding signal and the ratio=(S of non-specific signalsa+b/
BSA-Sb/BSA)÷Sb/BSA。
4, None group is processed as follows:
The reactant liquor BA20-STV of step 2 is passed through evanescent wave optical fiber sensing platform.During fiber laser arrays, first use
PBS rinses the optical fiber of the BSA-DTB modification of None group until baseline is steady；After baseline is steady, by reactant liquor BA20-
STV is liquid to be measured, opens peristaltic pump sample introduction 15s；Stop peristaltic pump afterwards, make liquid to be measured react 90s in reaction tank；Again open
Open peristaltic pump, be passed through eluent；Finally it is passed through PBS, steady to baseline, stop sampling simultaneously, preserve data (these data
For the mixed signal of specific binding signal Yu non-specific signals, by its named Sa+b/None)。
According to the method described above, BA20-STV replacing with the A20-STV of step 1, as comparison, other steps are the most constant,
Obtain non-specific signals, by its named Sb/None.Specific binding signal and the ratio=(S of non-specific signalsa+b/
None-Sb/None)÷Sb/None。
SSDNA suppression nucleic acid molecules to the closed process of sensing interface non-specific adsorption as shown in Figure 4, tie by above-mentioned experiment
Fruit is as shown in Figure 5.Result shows, on the optical fiber of None group, BSA group, A20 group and SSDNA group, nucleic acid molecules is specific binding
With the signal ratio respectively 1.0,5.8,9.5 and 40.2 of non-specific adsorption, using SSDNA to close strategy can be effectively by optical fiber
The signal of the specific binding and non-specific adsorption of nucleic acid molecules frequently rises to 40.20 times, and effect is significantly better than without closing
(None group), traditional BSA close (BSA group) and simple A oligomer closes (A20 group) strategy, use SSDNA to close plan
On optical fiber slightly, the signal of the specific binding and non-specific adsorption of nucleic acid molecules ratio is respectively without closing, traditional BSA envelope
Close and simple A oligomer closes tactful 40.20,6.93 and 4.23 times.
Claims (10)
1. suppress a nucleic acid molecules enclosure method to sensor interface non-specific adsorption, process including with salmon sperm dna
Sensor, it is achieved the closing of sensor.
Enclosure method the most according to claim 1, it is characterised in that: described salmon sperm dna is the salmon sperm dna of degeneration.
Enclosure method the most according to claim 1 and 2, it is characterised in that: described salmon sperm dna processes sensor and includes
Described sensor is processed with salmon sperm dna solution；Solute in described salmon sperm dna solution is salmon sperm dna.
Enclosure method the most according to claim 3, it is characterised in that: the solvent of described salmon sperm dna solution is PBS buffering
Liquid；
Described PBS is made up of solvent and solute, and described solvent is water, and described solute and concentration thereof are 0.1M phosphoric acid hydrogen two
Sodium and sodium dihydrogen phosphate and 0.15M sodium chloride, pH is 7.4.
5. according to the enclosure method described in claim 3 or 4, it is characterised in that: described salmon sperm dna is at described salmon sperm dna
Concentration in solution is 0.01mg/mL-10mg/mL.
6. according to described enclosure method arbitrary in claim 1-5, it is characterised in that: described salmon sperm dna processes sensing
The time of device or the described salmon sperm dna solution described sensor of process is 2-5 hour.
7. according to described enclosure method arbitrary in claim 1-6, it is characterised in that: described salmon sperm dna processes sensing
Device or described salmon sperm dna solution process described sensor all to be carried out at 30-37 DEG C.
8. according to described enclosure method arbitrary in claim 1-7, it is characterised in that: described sensor be biosensor or
Fibre Optical Sensor.
9. according to described enclosure method arbitrary in claim 1-7, it is characterised in that: described sensor is following a1) or a2)
Or a3) or a4):
A1) evanescent wave optical fiber sensor；
A2) nucleic acid sensor；
A3) optical fiber biosensor；
A4) evanescent wave optical fiber biosensor.
10. salmon sperm dna is suppressing nucleic acid molecules to the application in sensor interface non-specific adsorption.
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Application publication date: 20161207 |