CN102321759A - Fluorescence method for detecting S1 nuclease and inhibitor thereof - Google Patents
Fluorescence method for detecting S1 nuclease and inhibitor thereof Download PDFInfo
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- CN102321759A CN102321759A CN201110245341A CN201110245341A CN102321759A CN 102321759 A CN102321759 A CN 102321759A CN 201110245341 A CN201110245341 A CN 201110245341A CN 201110245341 A CN201110245341 A CN 201110245341A CN 102321759 A CN102321759 A CN 102321759A
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Abstract
The invention is a fluorescence method for detecting S1 nuclease and inhibitors thereof by using oligonucleotide and graphene oxide. The method uses graphene oxide as a solid carrier, allows a dye-labelled oligonucleotide probe to adhere to the surface of graphene oxide by the interaction between graphene oxide and nucleic acid bases, and performs fluorescence quenching. Undegraded complementary strands can form a double-spiral structure with the probe by hybridization, and are detached from the surface of graphene oxide, and thus the system fluorescence is recovered; in the presence of S1 nuclease, the complementary strands are degraded into small base gragments, and can not form a double-spiral structure with the probe, and the probe still adheres to the surface of graphene oxide, and the fluorescence is in a quenching state; adenosine triphosphate can effectively inhibit the degradation effect of nuclease on the complementary strands, and thus the system fluorescence is recovered. Therefore, by detecting the change of the system fluorescence intensity, the detection of S1 nuclease and inhibitors thereof can be realized. The method of the invention is simple and rapid, has high sensitivity and selectivity, and has important meaning with respect to nuclease detection.
Description
Technical field
The invention belongs to bio-sensing and analysis field, particularly a kind of fluorescence detection method that utilizes oligonucleotide and graphene oxide to detect nucleicacidase and suppressor factor thereof.
Background technology
Nucleicacidase is meant the enzyme of the phosphodiester bond that acts on nucleic acid.According to the different classification of substrate, can be divided into DNA enzyme and RNA enzyme; Different according to cutting part, can be divided into endonuclease, restriction enzyme and exonuclease again.Biological intravital nucleicacidase is responsible for the degraded of the inside and outside catalytic nucleic acid of cell, and vital process is played important effect.The purposes of nucleicacidase comprises that the important gene such as shearing, modification and degraded behind synthetic and the reparation and the rna transcription of participating in DNA duplicate and the genetic expression process; Be responsible for removing nucleic acid unnecessary, that 26S Proteasome Structure and Function is unusual, also can remove the exogenous nucleic acid of invading cell simultaneously; Nucleic acid in Digestive system in the degraded food is in order to absorption; It is the important tool enzyme in the extracorporeal recombinant DNA technology.
Because the significance of nucleicacidase physiological function receives the increasing concern of scientists in recent years, therefore press for quick, easy, the highly sensitive method of searching and detect nucleicacidase.Thereupon, the test material of nucleicacidase and the development research of method are become a focus.The novel analytical procedure that is directed to the nucleicacidase detection is at present developed rapidly in fields such as life science and disease molecular diagnosis.To deepening continuously of nucleicacidase research, will to the exploration of life quintessence with make bigger contribution human future.
Graphene oxide is learned and electrical properties with its special mechanics, quantum in recent years, paid attention to by physics and material educational circles.Advantages such as excellent electric property, thermal conductivity are good because it has, good water solubility, specific surface area are big also provide new thinking for the detection of biomolecules.Along with the successful Application of SWCN in bio-sensing, graphene oxide also becomes the object of sensory field favor gradually at present.Because the special property of graphene oxide makes that testing process is simple to operate, reaction is quick, has significantly reduced the step of experiment like this, can improve the sensitivity and the specificity of detection significantly simultaneously.
Summary of the invention
Technical problem: the technical problem that the present invention will solve is the deficiency that exists to existing nucleicacidase detection method; A kind of fluorescent method that utilizes oligonucleotide and graphene oxide to detect S1 nucleicacidase and suppressor factor thereof is provided, and it has higher specificity and sensitivity.
Technical scheme: the present invention detects the fluorescent method of S1 nucleicacidase and suppressor factor thereof, it is characterized in that this method may further comprise the steps:
A. the oligonucleotide fluorescent probe with dye marker joins in the buffered soln, is excitation wavelength with 480nm, carries out fluoroscopic examination, writes down the fluorescent emission bands of a spectrum of this probe;
B. graphene oxide is joined in the above-mentioned solution, graphene oxide adsorbs the oligonucleotide probe of free rolled state as solid phase carrier, forms oligonucleotide/graphene oxide mixture, at this moment the oxidized Graphene quencher of fluorescence of dyestuff;
C. through the oligonucleotide complementary strand of S1 nucleicacidase cutting, join in oligonucleotide/graphene complex and react;
D. the oligonucleotide complementary strand forms little base fragment through the specificity Degradation of S1 nucleicacidase; Can't form double-spiral structure with the oligonucleotide chain probe on graphene oxide surface; Thereby oligonucleotide probe can not break away from the surface of graphene oxide, and the fluorescent signal of dyestuff can not recover; The degree of recovering according to fluorescent signal realizes the qualitative and detection by quantitative to the S1 nucleicacidase;
E. the oligonucleotide complementary strand through the effect of S1 nuclease degradation, adds in oligonucleotide/graphene oxide mixture and reacts in the presence of the suppressor factor atp;
F. the oligonucleotide complementary strand can not formed double-spiral structure with oligonucleotide probe by the S1 nuclease degradation in the presence of the suppressor factor atp, and oligonucleotide probe breaks away from the surface of graphene oxide, and the fluorescent signal of dyestuff is able to recover; The degree of recovering according to fluorescent signal realizes the qualitative and detection by quantitative to the suppressor factor atp.
Wherein:
The described oligonucleotide fluorescent probe with dye marker of step a) joins in the buffered soln, detects with spectrophotofluorometer, and used dyestuff is a resorcinolphthalein; Dye marker is at 5 ' end of oligonucleotide probe; The oligonucleotide probe based composition is: 5 '-FAM-ATCTTGACTATGTGGGTGCT-3 '; Used buffered soln is 20mM Tris-HCl, 100mM NaCl, 5mM KCl, 5mM MgCl
2, pH=7.4; Used fluorescence detection method is: probe solution is joined the scanning of carrying out fluorescence emission spectrum in the 1.6mL buffered soln, and excitation wavelength is 480nm, and the emission wavelength sweep limit is 490-650nm.
The formation of the described oligonucleotide of step b)/graphene oxide mixture is to make oligonucleotide probe be adsorbed onto the graphene oxide surface through non-covalent interaction; After oligonucleotide is adsorbed onto the graphene oxide surface, the oxidized Graphene quencher of the fluorescence of dyestuff, join graphene oxide in the buffered soln that contains oligonucleotide probe after; Room temperature was placed 5 minutes; Carry out fluoroscopic examination then, excitation wavelength is 480nm, and sweep limit is 490-650nm.
Step c) joins in oligonucleotide/graphene complex and reacts through the oligonucleotide complementary strand of S1 nucleicacidase cutting, and used oligonucleotide complementary strand probe based composition is: 5 '-AGCACCCACATAGTCAAGAT-3 '; Adding behind the oligonucleotide complementary strand solution contain S 1 enzyme or other the isopyknic contrast solution fluorescent signal, to recover the used time be room temperature, about 25 minutes.
Described fluorescent signal detects with spectrophotofluorometer, and excitation wavelength is 480nm, and the emission wavelength sweep limit is 490-650nm.Described fluorescent probe is any one section single stranded oligonucleotide, and dye marker is at 5 ' end, or 3 ' end.Described dye marker is resorcinolphthalein or fluorescein isothiocyanate.
The described oligonucleotide complementary strand of step d) is through the effect of S1 nuclease degradation, be through the S1 nucleicacidase to single stranded DNA or the specific Degradation of RNA, form 5 ' phosphoric acid structure.
The S1 nucleicacidase is cut the used enzyme of process to the enzyme of oligonucleotide complementary strand, and to cut solution be 2mM CH
3COONa, 15mMNaCl, 0.1mMZnSO
4, pH=4.6; Enzyme Qie Wendu is 37 ℃, and the time is 30 minutes.
The described oligonucleotide complementary strand of step e) through the effect of S1 nuclease degradation, is that atp can suppress the specificity Degradation of S1 nucleicacidase to single stranded DNA or RNA cutting in the presence of the suppressor factor atp; When the suppressor factor atp existed, the S1 nucleicacidase was identical with above-mentioned enzyme tangent condition to the action condition of oligonucleotide complementary strand.
Beneficial effect:, can realize detection simultaneously to S1 nucleicacidase and suppressor factor atp thereof according to fluorescence detection method of the present invention.When containing other nucleicacidase and suppressor factor in the system, still can realize its high-sensitivity detection.Therefore, the inventive method is expected to detection for actual clinical sample amplifying nucleic acid enzyme provides a kind of easy, method fast.
Description of drawings
Fig. 1 detects the fundamental diagram of the fluorescence detection method of nucleicacidase and suppressor factor thereof for the present invention utilizes oligonucleotide and graphene oxide.
Fig. 2 detects without the fluoroscopic examination of the DNA chain of S1 enzyme liberating figure as a result for the present invention utilizes oligonucleotide and graphene oxide.A is an oligonucleotide probe; B is oligonucleotide probe+graphene oxide; C is oligonucleotide probe+complementary strand+graphene oxide; D is oligonucleotide probe+incomplementarity chain+graphene oxide.
Fig. 3 is a differential responses system fluoroscopic examination comparison diagram as a result in S1 enzyme and the suppressor factor testing process thereof.A is an oligonucleotide probe; B is oligonucleotide probe+graphene oxide; C is oligonucleotide probe+S1 nucleicacidase+complementary strand+graphene oxide; D is oligonucleotide probe+S1 nucleicacidase+atp+complementary strand+graphene oxide.
Embodiment
A kind of fluorescence detection method that utilizes oligonucleotide and graphene oxide to detect S1 nucleicacidase and suppressor factor thereof may further comprise the steps:
1) oligonucleotide probe with dye marker joins in the buffered soln, and solution is carried out fluoroscopic examination, writes down the fluorescent emission bands of a spectrum of this probe; This probe is one section single-chain nucleic acid of arbitrary sequence, and an end is marked with resorcinolphthalein.
2) graphene oxide is joined in the buffered soln as solid phase carrier; Because the graphene oxide surface has and can carry out non covalent bond bonded chemical group with oligonucleotide probe; Can make the oligonucleotide probe of dye marker be adsorbed onto the surface of graphene oxide; Thereby form oligonucleotide/graphene oxide mixture, taking place this moment efficiently between dyestuff and the graphene oxide, energy shifts the effective quencher of the oxidized Graphene of the fluorescence of dyestuff.
3) in oligonucleotide/graphene oxide mixture, add the oligonucleotide complementary strand and react, through base complementrity pairing effect, two strand Nucleotide hybridization form double-spiral structure.
4) the double-stranded formation of complementary strand mediation makes it break away from from the graphene oxide surface; Distance between dye groups and the graphene oxide is far away at this moment; Effective energy can not take place to be shifted; The fluorescent signal that is marked at the dyestuff of probe end is restored, and can detect stronger fluorescent signal.
5) when the oligonucleotide complementary strand after the S1 nuclease degradation forms little base fragment, join in oligonucleotide/graphene complex and react, the base fragment after the degraded can not with oligonucleotide chain formation double-spiral structure.
6) the oligonucleotide complementary strand can't form double-spiral structure with oligonucleotide owing to receive the effect of S1 nuclease degradation, thereby oligonucleotide probe can not break away from from the surface of graphene oxide, and the fluorescent signal of dyestuff can not recover.
7) when the oligonucleotide complementary strand in the presence of the suppressor factor atp after the effect of S1 nucleicacidase; Join in oligonucleotide/graphene complex and react; Therefore complementary strand can not formed double-spiral structure with oligonucleotide by the S1 nuclease degradation in the presence of the suppressor factor atp.
8) the oligonucleotide complementary strand since under the effect of suppressor factor atp not by the S1 nuclease degradation, can form double-spiral structure with oligonucleotide probe, thereby make oligonucleotide probe break away from the surface of graphene oxide, the fluorescent signal of dyestuff is able to recover.
According to the present invention, step 1) is: the oligonucleotide probe of dye marker is joined in the buffered soln, solution is carried out fluoroscopic examination; Wherein, oligonucleotide probe of the present invention is one section single-chain nucleic acid of arbitrary sequence.
The temperature that graphene oxide is joined in the buffered soln as solid phase carrier according to the present invention, step 2), dye marker oligonucleotide probe are adsorbed onto the graphene oxide surface can be a room temperature, preferably 25 ℃.
According to the present invention, step 2) it is very fast that the oligonucleotide probe described in is adsorbed onto the speed on graphene oxide surface, and the time of observing quenching of fluorescence is about 2 minutes.
According to the present invention, the temperature that the adding complementary strand reacts in oligonucleotide/graphene oxide mixture described in the step 3) can be a room temperature, preferably 25 ℃.Through base complementrity pairing effect, form double-spiral structure.
According to the present invention, after the oligonucleotide described in the step 4) and its complementary strand formed double-spiral structure, the time that breaks away from from the graphene oxide surface was about 25 minutes.
According to the present invention, the oligonucleotide complementary strand described in the step 5) can be 29 ℃～39 ℃ through the temperature of reaction of the Degradation of S1 nucleicacidase, preferably 37 ℃.
According to the present invention, the oligonucleotide complementary strand described in the step 5) is 30 minutes through the time of the Degradation of S1 nucleicacidase.
According to the present invention, the oligonucleotide complementary strand behind the process S1 nuclease degradation described in the step 6), adding the time of reacting in oligonucleotide/graphene complex is 25 minutes.
According to the present invention, described in the step 7) when the suppressor factor atp exists, in the oligonucleotide complementary strand, add the effect of S1 nucleicacidase, same step 5) of temperature of reaction and reaction times.
According to the present invention, described in the step 8) when the suppressor factor atp exists, through the oligonucleotide complementary strand after the effect of S1 nucleicacidase, add the time of reacting in oligonucleotide/graphene complex and be about 25 minutes.
Below in conjunction with accompanying drawing, further specify the present invention with the detection embodiment of S1 nucleicacidase.But the present invention is not limited in and detects nucleicacidase and suppressor factor thereof, does not also receive the restriction of other conditions among the embodiment.
The used instrument of fluoroscopic examination is fluorescent sub-photometer (RF-5301, a Japan).The fluorescence spectral measuring condition: xenon lamp excites, and exciting and launching slit width is 5.0nm and 10.0nm, and voltage is 950V, time of response 2S, and excitation wavelength is 480nm, emission wavelength sweep limit 490～650nm; Measure sample volume 1.60mL with the 3mL quartz colorimetric utensil; Room temperature.
Used graphene oxide is according to document (W.S.Hummers, R.E.Offeman, J.Am.Chem.Soc.1958,80,1339-1339) the method synthetic of middle Hummers in the embodiment of the invention.
Used oligonucleotide probe and complementary strand be all available from Shanghai biotechnology ltd in the embodiment of the invention, and sequence is respectively (5 '-FAM-ATCTTGACTATGTGGGTGCT-3 ') and (5 '-AGCACCCACATAGTCAAGAT-3 '); S1 nucleicacidase and atp are available from Shanghai biotechnology ltd; Enzyme is cut the liquid composition: (2mM CH
3COONa, 15mM NaCl, 0.1mMZnSO
4, pH 4.6; Used buffered soln composition is: 20mM Tris-HCl, 100mM NaCl, 5mMKCl, 5mM MgCl
2, pH 7.4.
Embodiment 1 preparation oligonucleotide/graphene oxide mixture
With 1 μ L concentration is that the fluorescein-labeled few nuclear nucleotide probe of 2 μ M joins in the Tris-HCl buffered soln of 1mL, adds 0.6mL water again and carries out fluoroscopic examination; And then to add 1.8 μ L concentration be the graphene oxide solution of 1mg/mL, and this probe can be adsorbed onto the graphene oxide surface rapidly, and with the signal quencher of resorcinolphthalein.
The detection of embodiment 2 oligonucleotide complementary strands
Detect 1: with 1 μ L concentration is that the oligonucleotide complementary strand of 30 μ M joins in oligonucleotide/graphene oxide mixture; Complementary strand and oligonucleotide probe form double-spiral structure; Break away from from the graphene oxide surface; (excitation wavelength 480nm, emission wavelength 490～650nm) can be observed the recovery phenomenon of fluorescence to carry out fluoroscopic examination.Be that the incomplementarity chain of 50 μ M adds as contrasting with 1 μ L concentration simultaneously, other condition is identical.
The detection of embodiment 3 nucleicacidases and suppressor factor thereof
Detect 2: with 1 μ L concentration is that the fluorescein-labeled few nuclear nucleotide probe of 1.6 μ M joins in the Tris-HCl buffered soln of 1mL, and the water that adds 0.6mL again carries out fluoroscopic examination; And then to add 1.5 μ L concentration be the graphene oxide solution of 1mg/mL, and this probe can be adsorbed onto the graphene oxide surface rapidly, and with the signal quencher of resorcinolphthalein.
With 3uL concentration is that the S1 nucleicacidase of 0.01U and oligonucleotide complementary strand that 1 μ L concentration is 30 μ M join the 3uL enzyme and cut liquid (2mM CH
3COONa, 15mM NaCl, 0.1mM ZnSO
4, pH 4.6) in, 37 ℃ were reacted 30 minutes down; Again reacted mixed solution is joined in oligonucleotide/graphene oxide mixture, as contrast, other condition with detect in 1 identical.
Simultaneously, do following test as a comparison.
Detect 3: with 2.5 μ L concentration is that the atp of 150mM and oligonucleotide complementary strand that 1 μ L concentration is 30 μ M join the 3uL enzyme and cut that (pH 4.6,2mM CH in the liquid
3COONa, 15mM NaCl, 0.1mM ZnSO
4), adding 3uL concentration again is the S1 nucleicacidase of 0.01U, 37 ℃ were reacted 30 minutes down; Reacted mixed solution is joined in oligonucleotide/graphene oxide mixture, as contrast, other condition with detect in 1 identical.
The result:
The fluorescent signal of oligonucleotide complementary strand and incomplementarity chain recovery intensity is compared and is differed greatly in the detection 1.It is thus clear that the selectivity of experiment is very strong.
The oligonucleotide complementary strand is compared through effect of S1 nuclease degradation and the Degradation when atp exists in the detection 2 and 3, and fluorescence intensity differs greatly.It is thus clear that the present invention has the specificity of height to the detection of nucleicacidase and suppressor factor thereof.
Claims (10)
1. fluorescent method that detects S1 nucleicacidase and suppressor factor thereof is characterized in that this method may further comprise the steps:
A. the oligonucleotide fluorescent probe with dye marker joins in the buffered soln, is excitation wavelength with 480nm, carries out fluoroscopic examination, writes down the fluorescent emission bands of a spectrum of this probe;
B. graphene oxide is joined in the above-mentioned solution, graphene oxide adsorbs the oligonucleotide probe of free rolled state as solid phase carrier, forms oligonucleotide/graphene oxide mixture, at this moment the oxidized Graphene quencher of fluorescence of dyestuff;
C. through the oligonucleotide complementary strand of S1 nucleicacidase cutting, join in oligonucleotide/graphene complex and react;
D. the oligonucleotide complementary strand forms little base fragment through the specificity Degradation of S1 nucleicacidase; Can't form double-spiral structure with the oligonucleotide chain probe on graphene oxide surface; Thereby oligonucleotide probe can not break away from the surface of graphene oxide, and the fluorescent signal of dyestuff can not recover; The degree of recovering according to fluorescent signal realizes the qualitative and detection by quantitative to the S1 nucleicacidase;
E. the oligonucleotide complementary strand through the effect of S1 nuclease degradation, adds in oligonucleotide/graphene oxide mixture and reacts in the presence of the suppressor factor atp;
F. the oligonucleotide complementary strand can not formed double-spiral structure with oligonucleotide probe by the S1 nuclease degradation in the presence of the suppressor factor atp, and oligonucleotide probe breaks away from the surface of graphene oxide, and the fluorescent signal of dyestuff is able to recover; The degree of recovering according to fluorescent signal realizes the qualitative and detection by quantitative to the suppressor factor atp.
2. the fluorescent method of detection S1 nucleicacidase according to claim 1 and suppressor factor thereof; It is characterized in that the described oligonucleotide fluorescent probe with dye marker of step a) joins in the buffered soln; Detect with spectrophotofluorometer, used dyestuff is a resorcinolphthalein; Dye marker is at 5 ' end of oligonucleotide probe; The oligonucleotide probe based composition is: 5 '-FAM-ATCTTGACTATGTGGGTGCT-3 '; Used buffered soln is 20mMTris-HCl, 100mM NaCl, 5mM KCl, 5mM MgCl
2, pH=7.4; Used fluorescence detection method is: probe solution is joined the scanning of carrying out fluorescence emission spectrum in the 1.6mL buffered soln, and excitation wavelength is 480nm, and the emission wavelength sweep limit is 490-650nm.
3. the fluorescent method of detection S1 nucleicacidase according to claim 1 and suppressor factor thereof; It is characterized in that the formation of the described oligonucleotide of step b)/graphene oxide mixture, is to make oligonucleotide probe be adsorbed onto the graphene oxide surface through non-covalent interaction; After oligonucleotide is adsorbed onto the graphene oxide surface, the oxidized Graphene quencher of the fluorescence of dyestuff, join graphene oxide in the buffered soln that contains oligonucleotide probe after; Room temperature was placed 5 minutes; Carry out fluoroscopic examination then, excitation wavelength is 480nm, and sweep limit is 490-650nm.
4. the fluorescent method of detection S1 nucleicacidase according to claim 1 and suppressor factor thereof; It is characterized in that the oligonucleotide complementary strand of step c) through the cutting of S1 nucleicacidase; Join in oligonucleotide/graphene complex and react, used oligonucleotide complementary strand probe based composition is: 5 '-AGCACCCACATAGTCAAGAT-3 '; Adding behind the oligonucleotide complementary strand solution contain S 1 enzyme or other the isopyknic contrast solution fluorescent signal, to recover the used time be room temperature, about 25 minutes.
5. the fluorescent method of detection S1 nucleicacidase according to claim 1 and suppressor factor thereof is characterized in that described fluorescent signal detects with spectrophotofluorometer, and excitation wavelength is 480nm, and the emission wavelength sweep limit is 490-650nm.
6. the fluorescent method of detection S1 nucleicacidase according to claim 1 and suppressor factor thereof is characterized in that described fluorescent probe is any one section single stranded oligonucleotide, and dye marker is at 5 ' end, or 3 ' end.
7. the fluorescent method of detection S1 nucleicacidase according to claim 1 and suppressor factor thereof is characterized in that described dye marker is resorcinolphthalein or fluorescein isothiocyanate.
8. the fluorescent method of detection S1 nucleicacidase according to claim 1 and suppressor factor thereof; It is characterized in that the described oligonucleotide complementary strand of step d) is through the effect of S1 nuclease degradation; Be through the S1 nucleicacidase to single stranded DNA or the specific Degradation of RNA, form 5 ' phosphoric acid structure.
9. the fluorescent method of detection according to claim 8 S1 nucleicacidase and suppressor factor thereof, it is characterized in that the S1 nucleicacidase enzyme of oligonucleotide complementary strand being cut the used enzyme of process, to cut solution be 2mM CH
3COONa, 15mMNaCl, 0.1mM ZnSO
4, pH=4.6; Enzyme Qie Wendu is 37 ℃, and the time is 30 minutes.
10. the fluorescent method of detection S1 nucleicacidase according to claim 1 and suppressor factor thereof; It is characterized in that the described oligonucleotide complementary strand of step e) in the presence of the suppressor factor atp through the effect of S1 nuclease degradation, be that atp can suppress the specificity Degradation of S1 nucleicacidase to single stranded DNA or RNA cutting; When the suppressor factor atp existed, the S1 nucleicacidase was identical with above-mentioned enzyme tangent condition to the action condition of oligonucleotide complementary strand.
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| CN102788824A (en) * | 2012-06-06 | 2012-11-21 | 张治红 | Preparation method of DNA biosensor |
| CN102998292A (en) * | 2012-12-13 | 2013-03-27 | 湖北中烟工业有限责任公司 | Fluorescence detection method for detecting ***e by use of oligonucleotide and graphene oxide |
| WO2014207515A1 (en) | 2013-06-25 | 2014-12-31 | Tubitak (Turkiye Bilimsel Ve Teknolojik Arastirma Kurumu) | Fluorescence method for detecting nuclease activity |
| CN107557431A (en) * | 2017-07-18 | 2018-01-09 | 天津大学 | A kind of kit of Visual retrieval S1 nucleases |
| WO2018167666A1 (en) | 2017-03-14 | 2018-09-20 | Tubitak | Method for rapid identification of microorganisms producing nuclease enzymes |
| CN111665351A (en) * | 2020-06-20 | 2020-09-15 | 江南大学 | Method for quickly and specifically determining RNA content |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102788824A (en) * | 2012-06-06 | 2012-11-21 | 张治红 | Preparation method of DNA biosensor |
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| WO2014207515A1 (en) | 2013-06-25 | 2014-12-31 | Tubitak (Turkiye Bilimsel Ve Teknolojik Arastirma Kurumu) | Fluorescence method for detecting nuclease activity |
| WO2018167666A1 (en) | 2017-03-14 | 2018-09-20 | Tubitak | Method for rapid identification of microorganisms producing nuclease enzymes |
| CN107557431A (en) * | 2017-07-18 | 2018-01-09 | 天津大学 | A kind of kit of Visual retrieval S1 nucleases |
| CN111665351A (en) * | 2020-06-20 | 2020-09-15 | 江南大学 | Method for quickly and specifically determining RNA content |
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