CN106872541A - A kind of probe system amplified for bio-sensor signal and its application - Google Patents
A kind of probe system amplified for bio-sensor signal and its application Download PDFInfo
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Abstract
The present invention is applied to electrochemica biological sensor field, there is provided a kind of probe system amplified for bio-sensor signal, the signal structure for amplifying being connected including joint probe and with the joint probe, wherein, the joint probe includes the second area that 3 ' ends are connected with the complementary first area for combining of target gene and with the signal structure for amplifying, the second area includes 24 identical sequence units, and base intervals are provided between the sequence units；The signal structure for amplifying includes 24 independent signal amplification units, the signal amplification unit is made up of signal probe with the series connection label for being connected to the end of signal probe 5 ', the signal probe is connected with sequence units complementation, and the series connection label is made up of the label of 24 series connection.Present invention also offers the detection method that a kind of electrochemical signals based on DNA/RNA hybridization concatenated probes amplify.
Description
Technical field
Put for bio-sensor signal the invention belongs to electrochemica biological sensor field, more particularly to one kind
Big probe system and its application.
Background technology
Hypersensitive nucleic acid detection technique should in the field such as the detection such as virus, pathogen, genopathy and its diagnosis
With extensive.Such as fluorescent quantitative PCR technique, rolling circle amplification (RCA) trace dna detection technique are based on
PCR and fluorescence signal detection method reach trace dna and quantify or detect, but these detection sides
Method has that operating procedure is complicated, optical signalling probe is expensive, need particular detection.Electrochemistry
Sensing technology is always most widely used biomedical and chemical analysis detection platform, with simple, portable
The advantages such as formula, low cost, high detection sensitivity.New electrochemica biological sensor is in bedside care diagnostic
(Point-Of-Care), the medical domain such as minimally invasive detection, antibiotic usage guidance has huge application prospect.
Traditional electrochemical nucleic acid detection sensor is mainly based on " sandwich " basic structure detection mode,
I.e. one end of target gene (TP) hybridizes with the capture probe (CP) containing its homologous complementary sequence, target gene
The other end and signal probe Complementary hybridization, by detect electrochemical parameter before and after hybridization (electric current, potential,
Conductance, impedance, electric capacity) change detect target dna.Traditional " sandwich " electrochemical method, one
One target dna of individual signal probe (DP) correspondence, when target dna content is too low, sensing detection method
The reasons such as the limitation of sensitivity itself, can cause background signal to be difficult to distinguish with faint hybridization signal.In disease
The fields such as disease early diagnosis, minimally invasive detection, food security, need highly sensitive biology sensor badly.Current DNA
Electrochemistry amplification detecting process mainly have two classes：Target is amplified and signal amplifies.Target scale-up version DNA
If sensor main is by Progress of Nucleic Acid Amplification Technologies, target DNA concentration to detection level is improved, realized
Trace DNA analysis, but the method needs the operating procedure of the expensive instrument and equipment of outfit and complexity, easily
The problems such as there is false positive.
The content of the invention
It is an object of the invention to provide a kind of probe system amplified for bio-sensor signal, it is intended to solve
The problem that certainly existing biology sensor sensitivity is low, complex operation or instrument and equipment are expensive.
Another object of the present invention is to provide a kind of electrochemistry letter based on DNA/RNA hybridization concatenated probes
Number amplify detection method.
The present invention is achieved in that a kind of probe system amplified for bio-sensor signal, including connects
Head probe and the signal structure for amplifying being connected with the joint probe,
Wherein, the joint probe include 3 ' the ends first area for combining complementary with target gene and with the signal
The second area of structure for amplifying connection, the second area includes 2-4 identical sequence units, the sequence
Base intervals are provided between column unit；
The signal structure for amplifying includes the independent signal amplification units of 2-4, the signal amplification unit by
Signal probe and be connected to the series connection label composition at the signal probe 5 ' end, the signal probe with it is described
The complementary connection of sequence units, the series connection label is made up of the label of 2-4 series connection.
And, the detection method that a kind of electrochemical signals based on DNA/RNA hybridization concatenated probes amplify,
Comprise the following steps：
Design provides capture probe, probe tip and the signal structure for amplifying of testing sample；
Electrode group is provided, the electrode group includes working electrode, auxiliary electrode and reference electrode, by the electricity
Pole carries out activation process, and the working electrode is carried out after the capture probe is fixed on the working electrode
Mercaptoethanol is closed, and obtains the DNA/RNA detection chips of capture probe specific recognition target gene；
The target gene of detected sample, the probe tip and the signal structure for amplifying are mixed to get detection
Liquid；
Liquid is detected in the DNA/RNA detection chips described in drop coating, treatment is incubated and is formed DNA/RNA
Hybridization concatenated probes chip, wherein, the DNA/RNA hybridization concatenated probes includes the claims 1-5
Any described probe system amplified for bio-sensor signal, in the joint probe described first
Region is complementary with the target gene to be combined；
Detect that the DNA/RNA hybridizes the current signal of concatenated probes.
Provided by the present invention for the probe system that bio-sensor signal is amplified, by the joint probe
The signal of the second area binding signal probe hybridization series connection label of (adapter probe, AP) amplifies
Structure, and the signal structure for amplifying includes 2-4 independent signal amplification unit, the series connection label
Including the 2-4 label of series connection, Signal averaging structure for amplifying is consequently formed, makes to connect on the signal probe
The electrochemical signals of the label for connecing amplify, so as to improve sensitivity.
The detection side that the electrochemical signals based on DNA/RNA hybridization concatenated probes that the present invention is provided amplify
Method, the method has the advantages that simple controllable, detection limit is low and signal is easy to collection, can quick detection it is to be measured
The concentration of sample, and sensitivity is high.
Brief description of the drawings
Fig. 1 is the probe system schematic diagram amplified for bio-sensor signal provided in an embodiment of the present invention；
Fig. 2 is the electrochemical signals that the use that the embodiment of the present invention 1 is provided is based on DNA hybridization concatenated probes
The e. coli dna concentration map that the detection method of amplification is measured；
Fig. 3 is tradition " sandwich formula " DNA sensing chips detection bacillus coli gene detection that comparative example is provided
The e. coli dna fragment concentrations gradient map that method is measured.
Specific embodiment
In order that the technical problem to be solved in the present invention, technical scheme and beneficial effect become more apparent, with
Lower combination drawings and Examples, the present invention will be described in further detail.It should be appreciated that described herein
Specific embodiment be only used to explain the present invention, be not intended to limit the present invention.
With reference to Fig. 1, a kind of probe system amplified for bio-sensor signal is the embodiment of the invention provides,
The signal structure for amplifying 4 being connected including joint probe 3 and with the joint probe 3, wherein, the joint
Probe 3 include 3 ' ends and the complementary first area 31 for combining of target gene 2 and with the signal structure for amplifying 4
The second area 32 of connection, the second area 32 includes 2-4 identical sequence units, the sequence
Base intervals are provided between unit；
The signal structure for amplifying 4 includes 2-4 independent signal amplification unit 41, and the signal amplifies single
Unit 41 is by signal probe 411 and 412 groups of the series connection label at the 5 ' ends for being connected to the signal probe 411
Into the signal probe 411 is connected with sequence units complementation, and the series connection label 412 is by 2-4
The label composition of individual series connection.
Specifically, in the embodiment of the present invention, the joint probe 3 is divided into two according to the difference of connecting object
Individual region, specifically, including the complementary first area 31 and 5 ' for combining of 3 ' ends and target gene 2 hold with it is described
The second area 32 of the connection of signal structure for amplifying 4.The embodiment of the present invention sets the signal structure for amplifying 4
At 5 ' ends of the joint probe 3, main reason is that the signal structure for amplifying 4 for being formed is carried with aftermentioned
The electrode 5 for arriving it is closer to the distance, it is easy to obtain electrochemical signals.
As the presently preferred embodiments, the base number of first area 31 described in the embodiment of the present invention is 15-25.
The suitable base number will not be because of the long increase design difficulty of length, or due to length in crossover process
It is long to cause top-cross；Also will not be not enough to carry the institute of its 5 ' section connection due to the too short adhesion of length not enough
State signal structure for amplifying 4.
The second area 32 is the region being connected with the signal structure for amplifying 4.The second area 32
2-4 identical sequence units are included according to the correspondence of the signal probe 411 that 2-4 complementary.As excellent
The sequence for selecting embodiment, the sequence units is：5'-CAACCTTCCCTCATTT-3'.
In order to weaken the space steric effect during signal probe series connection hybridization, between each sequence units
It is provided with base intervals.As the presently preferred embodiments, the base intervals are made up of two A bases.This is preferred
Base number and base type can ensure suitable space length between the sequence units, and can be with
Hybridization efficiency and accuracy when ensureing to hybridize.
The signal structure for amplifying 4 is the functional structure for amplifying biology sensor electrochemical signals, described
The number of signal probe amplifying unit in signal structure for amplifying 4, on the influence of the enlarging function of electrochemical signals compared with
Greatly, when the number of the signal probe amplifying unit 41 is in two or more, its signal amplifies can expire substantially
The test request of the various sample concentrations of foot, and the number of the signal probe amplifying unit 41 is more, the life
Thing sensor test sensitivity is relatively higher；When the number of the signal probe amplifying unit 41 is more than four
When, its signal amplification tends towards stability substantially.Accordingly, as preferred embodiment, the signal probe is put
The number of big unit 41 is preferably 2-4.Based on similar reason, the number of the label is preferably
2-4, when described and label number is excessive, easily forms larger steric hindrance, influence test knot
Really.
As the presently preferred embodiments, the sequence of the signal probe 411 is corresponded to：
5'-AAATGAGGGAAGGTTG-3'.The label be preferably ferrocene, fluorescein, biotin,
At least one in methylene blue.Further, because ferrocene is stable in properties not by oxygen concentration in environment
Change influence, the solubility in water is small, and electronics special delivery speed is fast, and oxidation-reduction potential is low, and the present invention is real
Apply and preferably use ferrocene as label in example.Using ferrocene as the label of the signal probe 411,
After being combined with electrolyte intermediate ion, oxidation-reduction potential changes, so as to realize the inspection of the fragment of target gene 2
Survey method.By the electricity of the label on the multiple described signal probe 411 that the joint probe 3 is captured
Signal averaging amplifies, and realizes that the detection sensitivity of bio-sensing chip detection target nucleic acid fragment is improved, up to fM
Below rank.
Used as a preferred embodiment, the second area 32 includes 4 identical sequence units, the letter
Number structure for amplifying 4 includes 4 independent signal amplification units 41, and the series connection label 412 is by 4 strings
The label composition of connection.
Additionally, the probe system for being used for bio-sensor signal amplification described in the embodiment of the present invention also includes target base
Because of 2, capture probe 1 and electrode 5, the capture probe 1, the first area 31 of the probe tip 3
Complementary with the target gene 1 respectively to combine, the one end of the capture probe 1 is connected with the electrode 5.Thus
The probe system amplified for bio-sensor signal for obtaining, by being fixed on described in gold electrode surfaces
Capture probe 1 and the complementary sequence hybridization of the target gene 2, Acquisition Detection sample target gene fragment, then lead to
Cross the first area 31 of the joint probe 3 and the hybridization of the target gene 2, the second area 32 with
The signal structure for amplifying 4 of the connection hybridization of signal probe polymer, forms multi signal superposition and amplifies polymer
Structure.
Further, as the presently preferred embodiments, in order that obtaining the signal connected in the probe tip 3
Structure for amplifying 4 and the electrode 5 it is closer to the distance, so as to preferably obtain electrochemical signals, the capture
0-12 bp is differed between the binding site that probe 1, probe tip 3 are engaged with the target gene 2 respectively.
Further, the electrode 5 is preferably gold electrode.
The probe system amplified for bio-sensor signal provided in an embodiment of the present invention, connects by described
The letter of the second area binding signal probe hybridization series connection label of head probe (adapter probe, AP)
Number structure for amplifying, and the signal structure for amplifying includes 2-4 independent signal amplification unit, the series connection
Label includes the 2-4 label of series connection, is consequently formed Signal averaging structure for amplifying, visits the signal
The electrochemical signals of the label connected on pin amplify, so as to improve sensitivity.
Accordingly, the embodiment of the present invention additionally provides a kind of electrification for hybridizing concatenated probes based on DNA/RNA
The detection method that signal amplifies is learned, is comprised the following steps：
S01. design provides capture probe, probe tip and the signal structure for amplifying of testing sample；
S02. electrode group is provided, the electrode group includes working electrode, auxiliary electrode and reference electrode, by institute
Stating electrode carries out activation process, is fixed on the working electrode after the capture probe to the working electrode
Mercaptoethanol closing is carried out, the DNA/RNA detection chips of capture probe specific recognition target gene are obtained；
S03. the target gene of detected sample, the probe tip and the signal structure for amplifying are mixed to get
Detection liquid；
S04. liquid is detected described in drop coating in the DNA/RNA detection chips, incubation treatment is formed
DNA/RNA hybridizes concatenated probes chip, wherein, the DNA/RNA hybridization concatenated probes includes above-mentioned use
In the probe system that bio-sensor signal is amplified, the first area and the target in the joint probe
Gene complementation is combined；
S05. detect that the DNA/RNA hybridizes the current signal of concatenated probes.
Specifically, in above-mentioned steps S01, the capture probe, probe tip and signal structure for amplifying set
Meter can be synthesized using computer software.The capture probe, probe tip and signal structure for amplifying are preferentially sieved
Choosing specificity height, sensitivity and reproducible optimal probe combinations.Wherein, the capture probe, described
The first area that probe tip is connected with target gene, should be designed according to the sequence of target gene.As
Specific embodiment, can first design synthesis target-gene sequence, and then by described in the target gene Series Design
The first area of probe tip.Wherein, the design of the synthesis target-gene sequence, can be by Genebank
The existing testing sample gene order of database and associated nucleic acid sequences reported in the literature are delivered both at home and abroad
Sequence alignment analysis are carried out, with testing sample gene without secondary structure and highly conserved section, according to drawing
Basic principle of the physical prospecting pin without mispairing, using software process quality.
In above-mentioned steps S02, the electrode group preferably uses film gold electrode.The method of the activation process
Can be realized using this area conventional method, concretely, electrode is immersed in the dilute sulfuric acid of 0.1M,
Electrochemical activation is carried out in the range of 0-1.2V, with ultrapure water electrode surface 10s, thoroughly cleaning electrode
The happiness acid of remained on surface, then nitrogen drying.
The method that the embodiment of the present invention fixes the capture probe on the working electrode is this area routine side
Method.As a specific embodiment, the μ L capture probe aqueous solution of drop coating 1,25 DEG C on the working electrode
Water-bath stands 2 hours, and the capture probe is fixed after terminating, with ultrapure water electrode surface 10 seconds, nitrogen
Air-blowing is done.The method of the mercaptoethanol closing, can also be realized, concretely using ability with conventional method：
The mercaptoethanol aqueous solution of the μ L1mM of drop coating 1,25 DEG C of water on the working electrode for securing the capture probe
Bath stands 2 hours, is dried up with nitrogen after ultrapure water, and capture probe/mercaptoethanol is formed in electrode surface
Composite bed.
In above-mentioned steps S03, the detection liquid should be included target gene, include the letter of signal probe
Number structure for amplifying, the mixed solution of probe tip.
In above-mentioned steps S04, as the presently preferred embodiments, the temperature for being incubated treatment is 25-42 DEG C, when
Between be 10-60min.Further, the temperature for being incubated treatment is 35 DEG C, and the time is 15min.This hair
In bright embodiment detection method, due to foring the probe system containing signal structure for amplifying, detection sensitivity
Improve, hybridization time is substantially shorter.
In above-mentioned steps S05, be electrolysed for DNA/RNA hybridization concatenated probes chips after terminating by hybridization
Liquid washs 10s, and the electrolyte can be using the NaClO of 1mol/L4(pH7.4).Then will be described
DNA/RNA hybridization concatenated probes chips are immersed in electrolyte, particularly fresh electrolyte, detect institute
State the common group of current signal of electrode.As the presently preferred embodiments, the DNA/RNA hybridization concatenated probes is detected
Current signal realized using alternating voltammetry or differential pulse voltammetry.Specifically, when using exchange volt-ampere
When method detects the current signal of the working electrode, voltage range is that voltage range is -0.3~0.4V, and frequency is
100-2000Hz, sweep speed is 0.01-0.05V/s.
The inspection that electrochemical signals based on DNA/RNA hybridization concatenated probes provided in an embodiment of the present invention amplify
Survey method, the method has the advantages that simple controllable, detection limit is low and signal is easy to collection, can quick detection
The concentration of testing sample, and sensitivity is high.
Illustrated with reference to specific embodiment.
The detection method detection large intestine that embodiment 1, the electrochemical signals based on DNA hybridization concatenated probes amplify
Bacillus gene, comprises the following steps：
S11. the design of probe：By to the Escherichia coli in the existing bacterial genomes of Genebank databases
16srRNA gene orders and associated nucleic acid sequences reported in the literature are delivered and have carried out sequence alignment both at home and abroad
Analysis, the conservative fragments with Escherichia coli 16srRNA gene orders as target gene, selection without secondary structure and
Highly conserved section, according to basic principle of the primed probe without mispairing, using software, engineer's capture
Probe, target-gene sequence, signal probe, joint probe.Detected respectively with multigroup probe of above-mentioned design
The synthesis target-gene sequence stated, through repetition test, filters out specificity, sensitivity and reproducible optimal
Probe combinations：CP, AP, DP1, the sequence of described CP, AP, DP1 are as follows respectively：
AP sequences：
5'-CAACCTTCCCTCATTTAACAACCTTCCCTCATTTAACAACCTTCCC
TCATTTAACAACCTTCCCTCATTTAACTTTACTCCCTTCC-3'。
CP sequences：
5'-S-S-C6-TCCCCGCTGAAAGTA CTTTACAACCCGAAGGCCTT-3'。
TP sequences：
5'-TGTATGAAGAAGGCCTTCGG
GTTGTAAAGTACTTTCAGCGGGGAGGAAGGGAGTAAAGTTAATACCTTT
GCTCATTGACGTTACCCGCAGAAGAAGCACC-3'。
DP1：
5'-FcFcFcFc-AAATGAGGGAAGGTTG-3', its 5'- end is labeled as 4 continuous two cyclopentadienyls
Iron (Fc).
It is prepared by S12.DNA chips：It is membrane electrode, working electrode, auxiliary electrode and reference that electrode is selected
Electrode is electrode.Three electrodes are immersed in the dilute sulfuric acid of 0.1M, are carried out in the range of 0V-1.2V
Electrochemical activation, with ultrapure water electrode surface 10 seconds, the diluted acid of thoroughly cleaning electrode surface residual, so
Nitrogen drying afterwards.CP is fixed：The μ L CP aqueous solution of drop coating 1 on the working electrode (s, it is small that 25 DEG C of water-baths stand 2
When, capture probe is fixed after terminating, with ultrapure water electrode surface 10 seconds, nitrogen drying.MCH is sealed
Close：The MCH aqueous solution of the μ L1mM of drop coating 1 on modified electrode, 25 DEG C of water-baths stand 2 hours, with super
Nitrogen drying after pure water rinsing, electrode surface forms CP/ mercaptoethanol composite beds, obtains specific recognition target
The DNA detection chips of gene.
It is prepared by S13 detection liquid：Detection liquid is the bacillus coli gene fragment (TP) comprising various concentrations, letter
Number probe (DP1), 6 × SSC (pH7.4) hybridization solution of joint probe (AP).The concentration of TP include 1pM,
100fM、10fM、1fM、100aM、0aM.The concentration of DP and AP is respectively 1 μM.
S14. hybridization reaction：The drop coating 1ul detections liquid on the modified DNA/ mercaptoethanol chips of CP,
35 DEG C are incubated 15 minutes.
S15. electrical signal detection：After hybridization terminates, by DNA/ mercaptoethanols chip electrolyte (1M
NaClO4, pH7.4) wash 10 seconds.Then electrode is immersed in fresh electrolyte, is lied prostrate using exchange
Peace method (ACV) detects the current signal of working electrode, and voltage range is -0.3~0.4V, and frequency is 250Hz,
Ac potential is 25mV, and sweep speed is 20mV/s.
Comparative example, target gene is detected using traditional " sandwich formula " DNA sensing chips
The design of probe：CP, AP are identical with above-mentioned S11, and the sequence of signal probe DP2 is
5'-Fc-AACTTTACTCCCTTCC-3', its 5'- end is labeled as 1 ferrocene (Fc).
It is prepared by detection liquid：Detection liquid is the bacillus coli gene fragment (TP) comprising various concentrations, signal spy
The bulk crossing liquid of pin (DP2).The concentration of TP includes 10pM, 1pM, 100fM, 10fM, 0pM.
The concentration of DP and AP is respectively 1 μM.
Hybridization reaction：The drop coating 1ul detections liquid on DNA/MCH-SAM chips.35 DEG C are incubated 15 minutes.
Electrical signal detection：Hybridization terminate after, by DNA/MCH-SAM chips with electrolyte (1M NaClO4,
PH7.4) wash 10 seconds, then electrode is immersed in fresh electrolyte, detected using alternating voltammetry
Electrode current signal, voltage range is -0.3~0.4V, and frequency is 250Hz, and ac potential is 25mV, is swept
Speed is retouched for 20mV/s.
The detection side that the electrochemical signals for being based on DNA hybridization concatenated probes using the embodiment of the present invention 1 amplify
The e. coli dna concentration map that method is measured is as shown in Figure 2.Traditional " sandwich formula " DNA sensing chips inspection
The e. coli dna fragment concentrations gradient that survey bacillus coli gene detection method is measured is as shown in Figure 3.By
Figure is visible, the inspection that the electrochemical signals based on DNA hybridization concatenated probes provided in an embodiment of the present invention amplify
Survey method has sensitivity higher, can obtain more accurately result.
Presently preferred embodiments of the present invention is the foregoing is only, is not intended to limit the invention, it is all in this hair
Any modification, equivalent and improvement made within bright spirit and principle etc., should be included in the present invention
Protection domain within.
Claims (10)
1. it is a kind of for bio-sensor signal amplify probe system, it is characterised in that including joint probe
And the signal structure for amplifying being connected with the joint probe,
Wherein, the joint probe include 3 ' the ends first area for combining complementary with target gene and with the signal
The second area of structure for amplifying connection, the second area includes 2-4 identical sequence units, the sequence
Base intervals are provided between column unit；
The signal structure for amplifying includes the independent signal amplification units of 2-4, the signal amplification unit by
Signal probe and be connected to the series connection label composition at the signal probe 5 ' end, the signal probe with it is described
The complementary connection of sequence units, the series connection label is made up of the label of 2-4 series connection.
It is 2. as claimed in claim 1 to be used for the probe system that bio-sensor signal is amplified, it is characterised in that
The sequence of the sequence units is：5'-CAACCTTCCCTCATTT-3'；Or
The sequence of the signal probe is：5'-AAATGAGGGAAGGTTG-3'.
It is 3. as claimed in claim 1 to be used for the probe system that bio-sensor signal is amplified, it is characterised in that
The base intervals are made up of two A bases.
4. the probe system amplified for bio-sensor signal as described in claim 1-3 is any, it is special
Levy and be, the label is at least one in ferrocene, fluorescein, biotin, methylene blue.
5. the probe system amplified for bio-sensor signal as described in claim 1-3 is any, it is special
Levy and be, also including target gene, capture probe and electrode, the capture probe, the of the probe tip
One region is complementary with the target gene respectively to be combined, and described capture probe one end is connected with the electrode.
It is 6. as claimed in claim 5 to be used for the probe system that bio-sensor signal is amplified, it is characterised in that
0-12 is differed between the binding site that the capture probe, probe tip are engaged with the target gene respectively
bp。
It is 7. as claimed in claim 5 to be used for the probe system that bio-sensor signal is amplified, it is characterised in that
The second area includes 4 identical sequence units, and the signal structure for amplifying includes 4 independent letters
Number amplifying unit, the series connection label is made up of the labels of 4 series connection.
8. the detection method that a kind of electrochemical signals based on DNA/RNA hybridization concatenated probes amplify, including
Following steps：
Design provides capture probe, probe tip and the signal structure for amplifying of testing sample；
Electrode group is provided, the electrode group includes working electrode, auxiliary electrode and reference electrode, by the electricity
Pole carries out activation process, and the working electrode is carried out after the capture probe is fixed on the working electrode
Mercaptoethanol is closed, and obtains the DNA/RNA detection chips of capture probe specific recognition target gene；
The target gene of detected sample, the probe tip and the signal structure for amplifying are mixed to get detection
Liquid；
Liquid is detected in the DNA/RNA detection chips described in drop coating, treatment is incubated and is formed DNA/RNA
Hybridization concatenated probes chip, wherein, the DNA/RNA hybridization concatenated probes includes the claims 1-7
Any described probe system amplified for bio-sensor signal, in the joint probe described first
Region is complementary with the target gene to be combined；
Detect that the DNA/RNA hybridizes the current signal of concatenated probes.
9. the electrochemical signals based on DNA/RNA hybridization concatenated probes as claimed in claim 8 amplify
Detection method, it is characterised in that the current signal of the detection DNA/RNA hybridization concatenated probes is using friendship
Stream voltammetry or differential pulse voltammetry realize
10. the electrochemical signals based on DNA/RNA hybridization concatenated probes as claimed in claim 8 amplify
Detection method, it is characterised in that the temperature of incubation treatment is 25-42 DEG C, and the time is 10-60min.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108048916A (en) * | 2017-12-20 | 2018-05-18 | 栾图 | Genetic chip for nucleic acid enriching extraction and preparation method thereof |
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