CN104101626B - Graphene modified HEMT is utilized to measure DNA hydridization method - Google Patents
Graphene modified HEMT is utilized to measure DNA hydridization method Download PDFInfo
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Abstract
The present invention relates to the use of graphene modified HEMT and measure the hydridization method of DNA, utilize graphene modified HEMT and fixing DNA probe, carry out the measurement of DNA hydridization, realizing the method utilizing graphene modified HEMT to measure DNA hydridization of the DNA hydridization detection of quick, sensitive and novel current-responsive pattern, the method builds HEMT first with molecular beam epitaxy；Followed by Graphene, DNA probe is fixed to surface, HEMT gate pole；Finally dropping target dna carries out hydridization measurement acquisition response current.Utilize this method can realize the detection of the DNA hydridization to actual sample.This method simplifies DNA probe and is fixed to the process of device surface, has given up the chemical process of complexity.This method obtains the DNA hydridization recognition mode of novelty.
Description
Technical field
The present invention relates to the transfer techniques of Graphene and the technique for fixing of DNA, utilize graphene modified HEMT (HEMT) and fixing DNA probe, carry out the measurement of DNA hydridization, it is achieved the DNA hydridization detection of quick, sensitive and novel current-responsive pattern.
Background technology
Realize rapid, the Sensitive Detection of DNA hydridization, play a very important role at aspects such as food safety detection, environmental monitoring, medical diagnosis on disease, criminal investigation investigation.At present the detection method of DNA mainly there are electrochemical method and photochemical method, although above method has been obtained for good testing result, but still there is the problem that detection efficiency is low and testing cost is expensive.Utilize field-effect transistor (FET) that DNA hydridization is detected, be a kind of effective means solving problem above.HEMT (HEMT), as a kind of special FET, has huge potential using value in terms of chemistry and biosensor.Owing to the two-dimensional electron gas in HEMT layer structure has high electron transfer capabilities, and its sensitive detection characteristic with excellence can be detected when the electron concentration on surface, HEMT gate pole has small change.Kang et al. just utilizes this characteristic of HEMT, and the DNA of thiol-based mistake is fixed on surface, HEMT gate pole, carries out DNA hydridization test (B.S.Kang, S.J.Pearton, J.J.Chen, etal, AppliedPhysicsLetters2006,89,122102.).DNA could be fixed to HEMT surface by complicated chemical process such as the job demand of Kang etc., and they do not mention the detection line by HEMT detection DNA hydridization and sensitivity in article.
Graphene is the new material of a kind of monolayer laminated structure being made up of carbon atom, and it is a kind of to be formed the flat film that hexangle type is honeycomb lattice, the two-dimensional material of only one of which carbon atom thickness by carbon atom with sp2 hybrid orbital.Graphene, with the structure of its uniqueness and excellent performance, has attracted substantial amounts of concern.In the numerous physics of Graphene, chemical property, high-specific surface area, easy functionalization, superior electron mobility and good biocompatibility so that it is in terms of biology sample detection, be provided with the highest potential using value.Graphene and derivant thereof are applied to enzyme bio-sensing and immune sensing detection has had numerous studies.Owing to DNA is combined with Graphene, DNA can be made to have the ability of the digestion resisting DNA enzymatic, there is many research to show, utilized Graphene and derivant thereof to build FET and DNA is detected, (R.Stine unmarked with DNA can be monitored in real time, J.T.Robinson, P.E.Sheehan, etal, Adv.Mater.2010,22,5297;Z.Yin, Q.He, X.Huang, Nanoscale2012,4,293.).Owing in actual sample, the content of DNA is often below 20fM(10-15Mol/L), FET-type DNA sensor needs higher sensitivity.
Summary of the invention
In order to solve the problems referred to above, the main object of the present invention is to realize rapid (in 5 minutes) of unmarked DNA sensor, super low concentration detection (less than 20fM), adapt to the demand of actual sample, accomplish to simplify the program that DNA is fixed on FET, and the method that the graphene modified HEMT obtaining the DNA hydridization recognition mode of novelty measures DNA hydridization simultaneously
The technical scheme is that and utilize graphene modified HEMT to measure DNA hydridization method
, specifically include following steps:
First, the structure of HEMT: utilize molecular beam epitaxy technique build HEMT layer structure, 580 DEG C, deposit GaAs layer 1 μm, Al0.26Ga0.7As layer 3nm, Si adulterate AlGaAs layer 22nm, and Si adulterates GaAs cap layers 5nm.Deposition Ni/AuGe/Ni/Au is as the source electrode of HEMT and drain electrode, wherein, Ni:50nm, AuGe:204nm, Ni:10nm, Au:50nm) as the source electrode of HEMT and drain electrode.Finally, utilize plasma enhanced chemical vapor deposition method by SiO2Insulating layer deposition is to device surface.Finally, with paraffin, HEMT is packaged, outside only keeping grid to be exposed to.
Secondly, DNA probe is fixed to HEMT gate pole: be 0.1-10mg/mL Graphene alcohol dispersion liquid by DNA probe solution with concentration, mix, its mixed proportion is DNA solution volume: graphene dispersing solution volume is 1:3-6, at temperature is 18-22 DEG C, mixed liquor is hatched 30-50 minute, appropriate artemia hatching solution drops in the HEMT gate that step 1 prepares extremely go up, carry out hatching and be dried at temperature is 0-4 DEG C, standby；Wherein, DNA probe solution is configured by phosphate buffer, and concentration is at 0.01-10 μM；
Finally, DNA hydridization is detected by graphene modified HEMT: the target dna of variable concentrations is added drop-wise to secure the HEMT gate pole of DNA probe, measures the curent change between HEMT source electrode and drain electrode, it is thus achieved that DNA hydridization electric current.
Advantages of the present invention or good effect:
Utilize this method can realize the detection of the DNA hydridization to actual sample.
This method simplifies DNA probe and is fixed to the process of device surface, has given up the chemical process of complexity.
This method obtains the DNA hydridization recognition mode of novelty.
Accompanying drawing explanation
Fig. 1 is the layer structure schematic diagram of HEMT.
Fig. 2 is DNA hydridization electric current " two steps " pattern；The curve synoptic diagram of any DNA response current " step " pattern.
Fig. 3 is the range of linearity and the Monitoring lower-cut curve synoptic diagram of graphene modified HEMT detection DNA hydridization.
In figure: 1.Ni/AuGe/Ni/Au electrode；2.SiO2Insulating barrier；3. grid；4.Si adulterates AlGaAs layer；5.GaAs layer；6. insulation GaAs sinks to the bottom；7.AlGaAs layer；8.Si adulterates AlGaAs cap layers.
Detailed description of the invention
Below in conjunction with specific embodiment, technical scheme is described further.
Embodiment 1:
Build the HEMT of 2mm × 5mm size.
With 1mg/mL, the Graphene alcohol dispersion liquid of 5 μ L and the DNA probe solution of 23 base pairs (0.1 μM, 20 μ L) at temperature is 18 DEG C, mixed liquor is hatched 30 minutes, mixed liquor is added drop-wise to surface, HEMT gate pole, 4 DEG C are dried and carry out DNA hydridization detection again in 12 hours.
The DNA sequence used in present embodiment is as follows:
DNA probe: 5 '-AAT-CAA-CTG-GGA-GAA-TGT-AAC-TG-3 '
Target dna: 5 '-CAG-TTA-CAT-TCT-CCC-AGT-TGA-TT-3 '
Any DNA: 5 '-ACC-TTC-CTC-CGC-AAT-ACT-CCC-3 '
The HEMT having secured DNA probe is carried out DNA hydridization detection, about 5 minutes i.e. available " two step pattern " DNA hydridization response currents.If dropping any DNA is to surface, HEMT gate pole, then can only obtain the response current of " step pattern ".The novel electric current recognition mode of graphene modified HEMT detection DNA hydridization is shown in Fig. 2.
At the target dna of HEMT surface dropping variable concentrations, obtain the range of linearity of DNA hydridization detection from 0.1fM to 0.1pM(10-12Mol/L), Monitoring lower-cut, up to 0.07fM, is shown in Fig. 3.
Embodiment 2:
Build the HEMT of 5mm × 5mm size.
With 0.5mg/mL, the Graphene alcohol dispersion liquid of 5 μ L and the DNA probe solution of 27 base pairs (0.1 μM, 25 μ L) at temperature is 20 DEG C, mixed liquor is hatched 40 minutes, mixed liquor is added drop-wise to surface, HEMT gate pole, 2 DEG C are dried and carry out DNA hydridization detection again in 12 hours.
The DNA sequence used in present embodiment is as follows:
DNA probe: 5 '-AAT-CAA-CTG-GGA-GAA-TGT-AAC-TGA-CCT-3 '
Target dna: 5 '-AGG-TCA-GTT-ACA-TTC-TCC-CAG-TTG-ATT-3 '
Any DNA: 5 '-ACC-TTC-CTC-CGC-AAT-ACT-CCC-ACT-CTG-3 '
The HEMT having secured DNA probe is carried out DNA hydridization detection, about 5 minutes i.e. available " two step pattern " DNA hydridization response currents.If dropping any DNA is to surface, HEMT gate pole, then can only obtain the response current of " step pattern ".
Embodiment 3:
Build the HEMT of 2mm × 2mm size.
With 1mg/mL, the Graphene alcohol dispersion liquid of 10 μ L and the DNA probe solution of 24 base pairs (0.2 μM, 30 μ L) at temperature is 22 DEG C, mixed liquor is hatched 50 minutes, mixed liquor is added drop-wise to surface, HEMT gate pole, 0 DEG C is dried and carries out DNA hydridization detection again in 12 hours.
The DNA sequence used in present embodiment is as follows:
DNA probe: 5 '-AAT-CAA-CTG-GGA-GAA-TGT-AAC-TGA-3 '
Target dna: 5 '-TCA-GTT-ACA-TTC-TCC-CAG-TTG-ATT-3 '
Any DNA: 5 '-TTC-CTC-CGC-AAT-ACT-CCC-ACT-CTG-3 '
The HEMT having secured DNA probe is carried out DNA hydridization detection, about 5 minutes i.e. available " two step pattern " DNA hydridization response currents.If dropping any DNA is to surface, HEMT gate pole, then can only obtain the response current of " step pattern ".
Claims (1)
1. utilize graphene modified HEMT to measure DNA hydridization method, it is characterised in that to specifically include following steps:
The structure of step 1. HEMT: at temperature 580 DEG C, molecular beam epitaxy deposition GaAs layer 1 μm, Al0.26Ga0.7As layer 3nm, Si adulterate AlGaAs layer 22nm, and Si adulterates GaAs cap layers 5nm；Deposition Ni/AuGe/Ni/Au is as the source electrode of HEMT and drain electrode, wherein, Ni:50nm, AuGe:204nm, Ni:10nm, Au:50nm；Utilize plasma enhanced chemical vapor deposition method by SiO2Insulating layer deposition is to device surface；With paraffin, HEMT is packaged, outside only keeping grid to be exposed to；
Step 2. DNA probe is fixed to HEMT grid: be 0.1-10mg/mL Graphene alcohol dispersion liquid by DNA probe solution with concentration, mix, its mixed proportion is DNA solution volume: Graphene alcohol dispersion liquid volume is 1:6-1:3, at temperature is 18-22 DEG C, mixed liquor is hatched 30-50 minute, subsequently, appropriate artemia hatching solution drops in the HEMT gate that step 1 prepares extremely go up, carry out hatching and be dried at temperature is 0-4 DEG C, standby；Wherein, DNA probe solution is configured by phosphate buffer, and concentration is at 0.01-10 μM；
DNA hydridization is detected by step 3. graphene modified HEMT: the target dna of variable concentrations is added drop-wise to secure the HEMT gate pole of DNA probe, measure the curent change between HEMT source electrode and drain electrode, it is thus achieved that DNA hydridization electric current.
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| US11215580B2 (en) | 2014-04-28 | 2022-01-04 | Cardea Bio, Inc. | System and method for DNA sequencing and blood chemistry analysis |
| US9765395B2 (en) | 2014-04-28 | 2017-09-19 | Nanomedical Diagnostics, Inc. | System and method for DNA sequencing and blood chemistry analysis |
| US20160054312A1 (en) | 2014-04-28 | 2016-02-25 | Nanomedical Diagnostics, Inc. | Chemically differentiated sensor array |
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