CN105154522A - Telomere sensor for detecting target DNAs and preparation method thereof - Google Patents
Telomere sensor for detecting target DNAs and preparation method thereof Download PDFInfo
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Abstract
The invention discloses a telomere sensor for detecting target DNAs and a preparation method thereof. The preparation method includes the steps: first, modifying a surface of each 0.2[mu]m carboxyl fluorescent microsphere with an amino DNA; second, hybridizing Alexa 488-DNA with the fluorescent microspheres/amino DNAs; third, forming a G-quadruplet. According to the method, the target DNAs are added to the fluorescent microspheres/amino DNAs/fluorescence modified DNAs, and reacting is carried out in the dark for 12h at a temperature below 25 DEG C. The telomere sensor is high in sensitivity and selectivity, a production method is simple, materials for use are easy to obtain, probe DNA sequences may be flexibly designed and modified according to differences between target DNA sequences, distances between the microspheres and Alexa 488 may also be adjusted by using amino DNAs different in length so as to obtain optimal detection effect. The telomere sensor is applicable to DNA detection and is highly worthy of application.
Description
Technical field
The invention belongs to biosensor, relate to a kind of for telomere sensor detecting target DNA and preparation method thereof.
Background technology
In recent years, biosensor is used to detect because its importance in the numerous areas such as molecular genetics has attracted extensive attention to DNA sequence dna, in order to improve sensitivity and the selectivity of detection, the multiple technologies such as optical technology, electronic conduction technology and surface plasma resonance technology obtain a large amount of uses.Wherein a kind of common methods is that enzyme, antibody or DNA probe are fixed to enzyme electrodes, DNA microarray etc. on the surface, because surface also exists nonspecific reaction, thus this method choice is poor, but by adding blocker, its specificity can be improved, make it play optimum performance.The method that another kind is used widely is homogeneous method, and reactant mixes by this method, reacts in the solution.In the past decade, the use of nanoparticle makes the signal expanded in equal phase medium, many homogeneous phase detection methods based on nanoparticle obtain significant progress, as silver amplification electro-detection technology, silver amplification Raman finger printing detection technique, magnetic particle DNA nanobarcode detection technique, conjugated polymers amplification fluorescent detection technique etc.In addition, have also appeared the new way using fluorescence dye, quantum dot and nano particle to detect DNA.
Many existing biosensors also exist.And telomere sensor of the present invention intends using the folded/expanded principle of G tetrad to build, its principle is, a kind of off-gauge secondary structure of the collapsible formation of rich GDNA sequence, i.e. G tetrad structure, this structure is very common in telomeric dna, after forming G tetrad structure, DNA sequence dna two ends distance significantly shortens, and when this structure is launched, distance increases, therefore the structural changes of G tetrad can be utilized, two kinds of fluorescence dyes are marked respectively as donor and acceptor at two ends, set up Fluorescence Resonance Energy transfering system, when G tetrad structure changes, supply, the spacing of acceptor changes, the corresponding change of FRET signal of system, utilize this principle, target DNA sequence with G tetrad complementary can be detected.
Summary of the invention
The object of the present invention is to provide a kind of for telomere sensor detecting target DNA and preparation method thereof, solve existing biosensor, as gold nano grain DNA sensor etc., the making of existence is complicated, sensitivity is lower, the problem of poor selectivity.
A kind ofly to comprise the following steps for telomere sensor detecting target DNA and preparation method thereof:
Step one, at the amino DNA of 0.2 μm of carboxyl fluorescent microsphere finishing, sets up the structure of fluorescent microsphere/amino DNA system;
Step 2, Alexa488-DNA and fluorescent microsphere/amino DNA hybridization, complete the structure of fluorescent microsphere/amino DNA/Alexa488-DNA system;
Step 3, the formation of G tetrad, completes the structure of telomere sensor.
Further, described step one concrete grammar:
Get 20 μ L carboxyl fluorescent microsphere suspension liquids, centrifuge washing, carboxyl fluorescent microsphere diameter 0.2 μm, structure is polymer wrapped crimson625/645 fluorescence dye, and finishing has carboxyl, and concentration is 2.3 × 10
12individual/mL; Again be dispersed to by microballoon in 100 μ LMES damping fluids, MES buffer concentration is 0.1M, pH5.75, adds the amino DNA of 35 μ L; Add 4 μ LEDAC solution, EDAC strength of solution is 10mg/mL, and solvent is MES damping fluid 0.1M, pH5.75; Sustained oscillation 1 hour at 25 DEG C, repeats four times; Centrifugal, with 140 μ LTEN buffer solution, TEN damping fluid is 1 × TEN damping fluid, pH7; Product fluorescent microsphere/amino DNA is dispersed to again in 140 μ LTEN damping fluids.
Further, described step 2 concrete grammar:
In fluorescent microsphere/amino DNA, add 18 μ LAlexa488-DNA, react 12 hours in dark surrounds at 25 DEG C, obtain product fluorescent microsphere/amino DNA/ fluorescent decoration DNA; Centrifugal, collect supernatant liquor.
Further, described step 3 concrete grammar:
Add part 34 μ LKCL solution, 18 μ LZnTCPP solution, KCl strength of solution is 0.588mM, ZnTCPP strength of solution is 100 μMs; React 8 hours in dark surrounds at 25 DEG C, obtain telomere sensor.
Further, amino DNA, Alexa488-DNA, target DNA concentration are all 100 μMs;
Sequence 3 ' the end of described amino DNA is marked with amino, and the sequence 3 ' end of Alexa488-DNA is marked with Alexa488 fluorescence dye;
The amino that the sequence 3 ' of described amino DNA is held is combined by the carboxyl of condensation reaction on microballoon, and Alexa488-DNA sequence is complementary with amino DNA, target DNA respectively.
Further, describedly a kind ofly specifically to comprise for telomere sensor detecting target DNA and preparation method thereof:
Step one, at 0.2 μm of carboxyl fluorescent microsphere finishing amino DNA, 5 '-ATGCTCGX-3 ', wherein X is amino:
Get 20 μ L2.3 × 10
12individual/mL carboxyl fluorescent microsphere suspension liquid, under 10000rpm centrifugal 10 minutes, with 0.1M, pH5.75MES buffer solution, is then again dispersed to microballoon in 100 μ LMES damping fluids, adds 35 μ L100 μM amino DNA; Add 4 μ L10mg/mLEDAC solution, sustained oscillation 1 hour at 25 DEG C, repeat four times; Centrifugal, with 140 μ LTEN (1 ×) buffer solution, product fluorescent microsphere/amino DNA is dispersed to again in 140 μ LTEN (1 ×) damping fluids;
Step 2, Alexa488-DNA and fluorescent microsphere/amino DNA hybridization:
In fluorescent microsphere/amino DNA, add 18 μ L100 μM Alexa488-DNA solution, react 12 hours in dark surrounds under 25 ° of C, obtain product fluorescent microsphere/amino DNA/ fluorescent decoration DNA; Centrifugal and collect supernatant liquor;
Step 3, the formation of G tetrad:
Add part 34 μ L0.588mMKCL solution, 18 μ L100 μM ZnTCPP solution, react 8 hours in dark surrounds under 25 ° of C, obtain telomere sensor.
Further, describedly a kind ofly specifically to comprise for telomere sensor detecting target DNA and preparation method thereof:
Step one, at 0.2 μm of carboxyl fluorescent microsphere finishing amino DNA, 5 '-ATGCTCGTTTTTX-3 ', wherein X is amino:
Get 20 μ L2.3 × 10
12individual/mL carboxyl fluorescent microsphere suspension liquid, under 10000rpm centrifugal 10 minutes, with 0.1M, pH5.75MES buffer solution, is then again dispersed to microballoon in 100 μ LMES damping fluids, adds 35 μ L100 μM amino DNA; Add 4 μ L10mg/mLEDAC solution, sustained oscillation 1 hour at 25 DEG C, repeat four times; Centrifugal, with 140 μ LTEN (1 ×) buffer solution, product fluorescent microsphere/amino DNA is dispersed to again in 140 μ LTEN (1 ×) damping fluids;
Step 2, Alexa488-DNA and fluorescent microsphere/amino DNA hybridization:
In fluorescent microsphere/amino DNA, add 18 μ L100 μM Alexa488-DNA solution, react 12 hours in dark surrounds under 25 ° of C, obtain product fluorescent microsphere/amino DNA/ fluorescent decoration DNA; Centrifugal and collect supernatant liquor;
Step 3, the formation of G tetrad:
Add part 34 μ L0.588mMKCL solution, 18 μ L100 μM ZnTCPP solution, react 8 hours in dark surrounds under 25 ° of C, obtain telomere sensor.
Further, describedly a kind ofly specifically to comprise for telomere sensor detecting target DNA and preparation method thereof:
Step one, at 0.2 μm of carboxyl fluorescent microsphere finishing amino DNA, 5 '-ATGCTCGTTTTTTTTTTX-3 ', wherein X is amino:
Get 20 μ L2.3 × 10
12individual/mL carboxyl fluorescent microsphere suspension liquid, under 10000rpm centrifugal 10 minutes, with 0.1M, pH5.75MES buffer solution, is then again dispersed to microballoon in 100 μ LMES damping fluids, adds 35 μ L100 μM amino DNA; Add 4 μ L10mg/mLEDAC solution, sustained oscillation 1 hour at 25 DEG C, repeat four times; Centrifugal, with 140 μ LTEN (1 ×) buffer solution, product fluorescent microsphere/amino DNA is dispersed to again in 140 μ LTEN (1 ×) damping fluids;
Step 2, Alexa488-DNA and fluorescent microsphere/amino DNA hybridization:
In fluorescent microsphere/amino DNA, add 18 μ L100 μM Alexa488-DNA solution, react 12 hours in dark surrounds under 25 ° of C, obtain product fluorescent microsphere/amino DNA/ fluorescent decoration DNA; Centrifugal and collect supernatant liquor;
Step 3, the formation of G tetrad:
Add part 34 μ L0.588mMKCL solution, 18 μ L100 μM ZnTCPP solution, react 8 hours in dark surrounds under 25 ° of C, obtain telomere sensor.
Another object of the embodiment of the present invention is the using method providing a kind of telomere sensor for detecting target DNA, the using method of the described telomere sensor for detecting target DNA is: in fluorescent microsphere/amino DNA/ fluorescent decoration DNA, add 18 μ L100 μM target DNAs, reacts 12 hours at 25 DEG C in dark surrounds.
Provided by the invention for telomere sensor detecting target DNA and preparation method thereof and using method, telomere sensor, highly sensitive, selectivity good, making method is simple, use material is easy to get, DNA probe sequence can be changed according to the different flexible design of target DNA sequence, and distance between microballoon and Alexa488 can be regulated, reaching optimum detection effect by using the amino DNA of different lengths.This Novel DNA sensor of the present invention can be used for DNA detection, has very large using value.
Accompanying drawing explanation
Fig. 1 is telomere sensor construction and the Cleaning Principle schematic diagram for detecting target DNA that the embodiment of the present invention provides;
Fig. 2 is telomere transducer production method and the use schematic flow sheet for detecting target DNA that the embodiment of the present invention provides;
Fig. 3 be the embodiment of the present invention provide for detecting the fluorescence spectrum figure in the telomere sensor preparation process of target DNA;
Fig. 4 is the fluorescence spectrum comparison diagram that telomere sensor that the embodiment of the present invention provides adds after target DNA and not complementary DNA;
Fig. 5 is the fluorescence spectrum figure of telomere sensor prepared by the amino DNA of the use different lengths that provides of the embodiment of the present invention.
Embodiment
Below in conjunction with embodiment, the present invention is described in detail.
As shown in Figure 1, the structure for the telomere sensor detecting target DNA that the embodiment of the present invention provides is specially: carboxyl fluorescent microsphere is connected with amino DNA, amino DNA and Alexa488-DNA is connected by base complementrity, and Alexa488-DNA forms G-tetrad structure near 3 ' end place;
As shown in Figure 2, a kind of of the embodiment of the present invention comprises the following steps for telomere sensor detecting target DNA and preparation method thereof:
Step one, at the amino DNA of 0.2 μm of carboxyl fluorescent microsphere finishing:
Get 20 μ L2.3 × 10
12individual/mL carboxyl fluorescent microsphere suspension liquid, under 10000rpm centrifugal 10 minutes, with 0.1M, pH5.75MES buffer solution, is then again dispersed to microballoon in 100 μ LMES damping fluids, adds 35 μ L100 μM amino DNA; Add 4 μ L10mg/mLEDAC solution, sustained oscillation 1 hour at 25 DEG C, repeat four times; Centrifugal, with 140 μ LTEN (1 ×) buffer solution, product fluorescent microsphere/amino DNA is dispersed to again in 140 μ LTEN (1 ×) damping fluids;
Step 2, Alexa488-DNA and fluorescent microsphere/amino DNA hybridization:
In fluorescent microsphere/amino DNA, add 18 μ L100 μM Alexa488-DNA solution, react 12 hours in dark surrounds under 25 ° of C, obtain product fluorescent microsphere/amino DNA/ fluorescent decoration DNA; Centrifugal and collect supernatant liquor;
Step 3, the formation of G tetrad:
Add part 34 μ L0.588mMKCL solution, 18 μ L100 μM ZnTCPP solution, react 8 hours in dark surrounds under 25 ° of C, obtain telomere sensor.
Step 4, the hybridization of sensor and target DNA:
In fluorescent microsphere/amino DNA/ fluorescent decoration DNA, add 18 μ L target DNAs, react 12 hours in dark surrounds at 25 DEG C.
Described carboxyl fluorescent microsphere diameter about 0.2 μm, structure is polymer wrapped crimson625/645 fluorescence dye, and finishing has carboxyl.
Described carboxyl fluorescent microsphere turbid liquid concentration is 2.3 × 10
12individual/mL.
Described MES buffer concentration is 0.1M, pH5.75.
Described amino DNA, Alexa488-DNA, target DNA concentration are all 100 μMs.
Described amino DNA sequence dna 3 ' end is marked with amino, and Alexa488-DNA sequence 3 ' end is marked with Alexa488 fluorescence dye.
The amino that described amino DNA sequence dna 3 ' is held is combined by the carboxyl of condensation reaction on microballoon, and Alexa488-DNA sequence A lexa488 is complementary with amino DNA, target DNA respectively.
Described EDAC strength of solution is 10mg/mL, and solvent is MES damping fluid (0.1M, pH5.75).
Described TEN damping fluid is 1 × TEN damping fluid, pH7.
Described KCl strength of solution is 0.588mM.
Described ZnTCPP strength of solution is 100 μMs.
Below in conjunction with specific embodiment, principle of the present invention is described in detail:
Embodiment 1:
A method of preparation and use for telomere sensor, its concrete technical scheme is as follows:
Step one, at the amino DNA of 0.2 μm of carboxyl fluorescent microsphere finishing (5 '-ATGCTCGX-3 ', wherein X be amino):
Get 20 μ L2.3 × 10
12individual/mL carboxyl fluorescent microsphere suspension liquid, under 10000rpm centrifugal 10 minutes, with 0.1M, pH5.75MES buffer solution, is then again dispersed to microballoon in 100 μ LMES damping fluids, adds 35 μ L100 μM amino DNA; Add 4 μ L10mg/mLEDAC solution, sustained oscillation 1 hour at 25 DEG C, repeat four times; Centrifugal, with 140 μ LTEN (1 ×) buffer solution, product fluorescent microsphere/amino DNA is dispersed to again in 140 μ LTEN (1 ×) damping fluids;
Step 2, Alexa488-DNA and fluorescent microsphere/amino DNA hybridization:
In fluorescent microsphere/amino DNA, add 18 μ L100 μM Alexa488-DNA solution, react 12 hours in dark surrounds under 25 ° of C, obtain product fluorescent microsphere/amino DNA/ fluorescent decoration DNA; Centrifugal and collect supernatant liquor;
Step 3, the formation of G tetrad:
Add part 34 μ L0.588mMKCL solution, 18 μ L100 μM ZnTCPP solution, react 8 hours in dark surrounds under 25 ° of C, obtain telomere sensor.
Using method: the hybridization of sensor and target DNA
In fluorescent microsphere/amino DNA/ fluorescent decoration DNA, add 18 μ L100 μM target DNAs, react 12 hours in dark surrounds under 25 ° of C.
Embodiment 2
A method of preparation and use for telomere sensor, its concrete technical scheme is as follows:
Step one, at the amino DNA of 0.2 μm of carboxyl fluorescent microsphere finishing (5 '-ATGCTCGTTTTTX-3 ', wherein X be amino):
Get 20 μ L2.3 × 10
12individual/mL carboxyl fluorescent microsphere suspension liquid, under 10000rpm centrifugal 10 minutes, with 0.1M, pH5.75MES buffer solution, is then again dispersed to microballoon in 100 μ LMES damping fluids, adds 35 μ L100 μM amino DNA; Add 4 μ L10mg/mLEDAC solution, sustained oscillation 1 hour at 25 DEG C, repeat four times; Centrifugal, with 140 μ LTEN (1 ×) buffer solution, product fluorescent microsphere/amino DNA is dispersed to again in 140 μ LTEN (1 ×) damping fluids;
Step 2, Alexa488-DNA and fluorescent microsphere/amino DNA hybridization:
In fluorescent microsphere/amino DNA, add 18 μ L100 μM Alexa488-DNA solution, react 12 hours in dark surrounds under 25 ° of C, obtain product fluorescent microsphere/amino DNA/ fluorescent decoration DNA; Centrifugal and collect supernatant liquor;
Step 3, the formation of G tetrad:
Add part 34 μ L0.588mMKCL solution, 18 μ L100 μM ZnTCPP solution, react 8 hours in dark surrounds under 25 ° of C, obtain telomere sensor.
Using method: the hybridization of sensor and target DNA:
In fluorescent microsphere/amino DNA/ fluorescent decoration DNA, add 18 μ L50 μM target DNAs, react 12 hours in dark surrounds under 25 ° of C.
Embodiment 3
A method of preparation and use for telomere sensor, its concrete technical scheme is as follows:
Step one, at the amino DNA of 0.2 μm of carboxyl fluorescent microsphere finishing (5 '-ATGCTCGTTTTTTTTTTX-3 ', wherein X be amino):
Get 20 μ L2.3 × 10
12individual/mL carboxyl fluorescent microsphere suspension liquid, under 10000rpm centrifugal 10 minutes, with 0.1M, pH5.75MES buffer solution, is then again dispersed to microballoon in 100 μ LMES damping fluids, adds 35 μ L100 μM amino DNA; Add 4 μ L10mg/mLEDAC solution, sustained oscillation 1 hour at 25 DEG C, repeat four times; Centrifugal, with 140 μ LTEN (1 ×) buffer solution, product fluorescent microsphere/amino DNA is dispersed to again in 140 μ LTEN (1 ×) damping fluids;
Step 2, Alexa488-DNA and fluorescent microsphere/amino DNA hybridization:
In fluorescent microsphere/amino DNA, add 18 μ L100 μM Alexa488-DNA solution, react 12 hours in dark surrounds under 25 ° of C, obtain product fluorescent microsphere/amino DNA/ fluorescent decoration DNA; Centrifugal and collect supernatant liquor;
Step 3, the formation of G tetrad:
Add part 34 μ L0.588mMKCL solution, 18 μ L100 μM ZnTCPP solution, react 8 hours in dark surrounds under 25 ° of C, obtain telomere sensor.
Using method: the hybridization of sensor and target DNA
In fluorescent microsphere/amino DNA/ fluorescent decoration DNA, add 10 μ L50 μM target DNAs, react 12 hours in dark surrounds under 25 ° of C.
By following comparing result, effect of the present invention is described in detail:
As shown in Figure 3, donor (Alexa488) fluorescence peak is at 535nm place, and acceptor (crimson625) fluorescence peak is at 647nm place.After adding ligand reaction, donor fluorescent weakens, and acceptor fluorescence strengthens, and FRET strengthens, and Distance Shortened between confession, acceptor is described, G tetrad structure is formed, and illustrates that telomere sensor is made.
As shown in Figure 4, add target DNA reaction in sensor after, donor fluorescence intensity obviously strengthens, and acceptor fluorescence intensity significantly reduces, and FRET signal significantly weakens; And in sensor, adding not complementary DNA, FRET signal, without too large change, can prove thus, and telomere sensor successfully can detect target DNA, and selectivity is good.
Five amino DNA sequence dnas are respectively from short to long as shown in Figure 5: 5 '-ATGCTCGX-3 ' (sequence numbering I), 5 '-ATGCTCGTTTTTX-3 ' (sequence numbering II), 5 '-ATGCTCGTTTTTTTTTTX-3 ' (sequence numbering III), 5 '-ATGCTCGTTTTTTTTTTTTTTTX-3 ' (sequence numbering IV), 5 '-ATGCTCGTTTTTTTTTTTTTTTTTTTTX-3 ' (sequence numbering V), wherein X represents amino.As seen from the figure, the telomere sensor FRET signal using the amino DNA of different lengths to prepare weakens gradually along with the growth of amino DNA sequence dna.
The above is only to better embodiment of the present invention, not any pro forma restriction is done to the present invention, every any simple modification done above embodiment according to technical spirit of the present invention, equivalent variations and modification, all belong in the scope of technical solution of the present invention.
Claims (7)
1. one kind for telomere sensor detecting target DNA and preparation method thereof, it is characterized in that according to following steps:
Step one, at the amino DNA of 0.2 μm of carboxyl fluorescent microsphere finishing;
Step 2, Alexa488-DNA and fluorescent microsphere/amino DNA hybridization;
Step 3, the formation of G tetrad.
2. a kind ofly as claimed in claim 1 to it is characterized in that for telomere sensor detecting target DNA and preparation method thereof, described step one concrete grammar:
Get 20 μ L carboxyl fluorescent microsphere suspension liquids, centrifuge washing, carboxyl fluorescent microsphere diameter 0.2 μm, structure is polymer wrapped crimson625/645 fluorescence dye, and finishing has carboxyl, and concentration is 2.3 × 10
12individual/mL; Again be dispersed to by microballoon in 100 μ LMES damping fluids, MES buffer concentration is 0.1M, pH5.75, adds the amino DNA of 35 μ L; Add 4 μ LEDAC solution, EDAC strength of solution is 10mg/mL, and solvent is MES damping fluid 0.1M, pH5.75; Sustained oscillation 1 hour at 25 DEG C, repeats four times; Centrifugal, with 140 μ LTEN buffer solution, TEN damping fluid is 1 × TEN damping fluid, pH7; Product fluorescent microsphere/amino DNA is dispersed to again in 140 μ LTEN damping fluids.
3. a kind ofly as claimed in claim 1 to it is characterized in that for telomere sensor detecting target DNA and preparation method thereof, described step 2 concrete grammar:
In fluorescent microsphere/amino DNA, add 18 μ LAlexa488-DNA, react 12 hours in dark surrounds at 25 DEG C, obtain product fluorescent microsphere/amino DNA/ fluorescent decoration DNA; Centrifugal, collect supernatant liquor.
4. if claim 1 one kind is for telomere sensor detecting target DNA and preparation method thereof, it is characterized in that, described step 3 concrete grammar:
Add part 34 μ LKCL solution, 18 μ LZnTCPP solution, KCl strength of solution is 0.588mM, ZnTCPP strength of solution is 100 μMs; React 8 hours in dark surrounds at 25 DEG C, obtain telomere sensor.
5. if claim 1 one kind is for telomere sensor detecting target DNA and preparation method thereof, it is characterized in that, amino DNA, Alexa488-DNA, target DNA concentration are all 100 μMs;
Sequence 3 ' the end of described amino DNA is marked with amino, and the sequence 3 ' end of Alexa488-DNA is marked with Alexa488 fluorescence dye;
The amino that the sequence 3 ' of described amino DNA is held is combined by the carboxyl of condensation reaction on microballoon, and Alexa488-DNA sequence A lexa488 is complementary with amino DNA, target DNA respectively.
6. if claim 1 one kind is for telomere sensor detecting target DNA and preparation method thereof, it is characterized in that, the preparation method of the described telomere sensor for detecting target DNA specifically comprises:
Step one, at 0.2 μm of carboxyl fluorescent microsphere finishing amino DNA, 5 '-ATGCTCGX-3 ', wherein X is amino:
Get 20 μ L2.3 × 10
12individual/mL carboxyl fluorescent microsphere suspension liquid, under 10000rpm centrifugal 10 minutes, with 0.1M, pH5.75MES buffer solution, is then again dispersed to microballoon in 100 μ LMES damping fluids, adds 35 μ L100 μM amino DNA; Add 4 μ L10mg/mLEDAC solution, sustained oscillation 1 hour at 25 DEG C, repeat four times; Centrifugal, with 140 μ LTEN (1 ×) buffer solution, product fluorescent microsphere/amino DNA is dispersed to again in 140 μ LTEN (1 ×) damping fluids;
Step 2, Alexa488-DNA and fluorescent microsphere/amino DNA hybridization:
In fluorescent microsphere/amino DNA, add 18 μ L100 μM Alexa488-DNA solution, react 12 hours in dark surrounds at 25 DEG C, obtain product fluorescent microsphere/amino DNA/ fluorescent decoration DNA; Centrifugal and collect supernatant liquor;
Step 3, the formation of G tetrad:
Add part 34 μ L0.588mMKCL solution, 18 μ L100 μM ZnTCPP solution, react 8 hours in dark surrounds at 25 DEG C, obtain telomere sensor.
7. according to claim 1 method prepare a kind of for telomere sensor detecting target DNA and preparation method thereof, it is characterized in that: in fluorescent microsphere/amino DNA/ fluorescent decoration DNA, add target DNA, at 25 DEG C in dark surrounds react 12 hours.
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| CN107058585A (en) * | 2017-06-07 | 2017-08-18 | 浙江殷欣生物技术有限公司 | A kind of detection method of nucleic acid hybridization |
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| CN107058585A (en) * | 2017-06-07 | 2017-08-18 | 浙江殷欣生物技术有限公司 | A kind of detection method of nucleic acid hybridization |
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